Recombinant poxvirus-cytomegalovirus, compositions and uses

ABSTRACT

Attenuated recombinant viruses containing DNA encoding an HCMV antigen, as well as methods and compositions employing the viruses, expression products therefrom, and antibodies generated from the viruses or expression products, are disclosed and claimed. The recombinant viruses can be NYVAC or ALVAC recombinant viruses. The recombinant viruses and gene products therefrom and antibodies generated by the viruses and gene products have several preventive, therapeutic and diagnostic uses. The DNA of the recombinant viruses can be used as probes or for generating PCR primers.

RELATED APPLICATIONS

This application is a continuation-in-part of application Ser. No. 08/471,014, filed Jun. 6, 1995, now abandoned, which in turn is a continuation-in-part of application Ser. No. 08/105,483, filed Aug. 13, 1993, now U.S. Pat. No. 5,494,807, which in turn is a continuation of application Ser. No. 07/847,951, filed Mar. 6, 1992, now abandoned, which in turn is a continuation-in-part of application Ser. No. 07/713,967, filed Jun. 11, 1991, now abandoned, which in turn is a continuation-in-part of application Ser. No. 07/666,056, filed Mar. 7, 1991, now abandoned. This application is also a continuation-in-part of U.S. application Ser. No. 08/124,668, filed Sep. 21, 1993, now U.S. Pat. No. 5,482,713 as a divisional of application Ser. No. 07/502,834, filed Apr. 4, 1990, now U.S. Pat. No. 5,338,683; application Ser. No. 07/502,834 was a continuation-in-part of application Ser. No. 07/394,488, filed Aug. 16, 1989, now abandoned, which was a continuation-in-part of application Ser. No. 07/339,004, filed Apr. 17, 1989, now abandoned; and, a continuation-in-part of application Ser. No. 07/090,209, filed Aug. 27, 1987, now abandoned, which was a division of application Ser. No. 06/622,135, filed Jun. 19, 1984, now U.S. Pat. No. 4,722,848, which was a continuation-in-part of application Ser. No. 06/446,824, filed Dec. 8, 1982, now U.S. Pat. No. 4,603,112, which was a continuation-in-part of U.S. application Ser. No. 06/334,456, filed Dec. 24, 1981, now U.S. Pat. No. 4,769,330. Each of the aforementioned and above-referenced applications and patents are hereby incorporated herein by reference.

FIELD OF THE INVENTION

The present invention relates to a modified poxvirus and to methods of making and using the same; for instance, a vaccinia virus or avipox (e.g. canarypox or fowlpox), e.g., modified recombinant poxvirus-cytomegalovirus (CMV), e.g., human cytomegalovirus (HCMV) such as an attenuated recombinant, especially a NYVAC or ALVAC CMV or HCMV recombinant. More in particular, the invention relates to improved vectors for the insertion and expression of foreign genes for use as safe immunization vehicles to elicit an immune response against CMV or HCMV virus. Thus, the invention relates to a recombinant poxvirus, which virus expresses gene products of CMV or HCMV and to immunogenic compositions which induce an immunological response against CMV or HCMV infections when administered to a host, or in vitro (e.g., ex vivo modalities) as well as to the products of expression of the poxvirus which by themselves are useful for eliciting an immune response e.g., raising antibodies, which antibodies are useful against CMV or HCMV infection, in either seropositive or seronegative individuals, or which expression products or antibodies elicited thereby, isolated from an animal or human or cell culture as the case may be, are useful for preparing a diagnostic kit, test or assay for the detection of the virus, or of infected cells, or, of the expression of the antigens or products in other systems. The isolated expression products are especially useful in kits, tests or assays for detection of antibodies in a system, host, serum or sample, or for generation of antibodies. The poxvirus recombinants preferably contain DNA coding for any or all of CMV or HCMVgB, gH, gL, pp150, pp65 and IE1, including recombinants expressing truncated versions of IE1; and, the recombinant poxvirus DNA is useful for probes for CMV or HCMV or for preparing PCR primers for detecting the presence or absence of CMV or HCMV or antigens thereof.

Several publications are referenced in this application. Full citation to these references is found at the end of the specification immediately preceding the claims or where the publication is mentioned; and each of these publications is hereby incorporated herein by reference.

BACKGROUND OF THE INVENTION

Vaccinia virus and more recently other poxviruses have been used for the insertion and expression of foreign genes. The basic technique of inserting foreign genes into live infectious poxvirus involves recombination between pox DNA sequences flanking a foreign genetic element in a donor plasmid and homologous sequences present in the rescuing poxvirus (Piccini et al., 1987).

Specifically, the recombinant poxviruses are constructed in two steps known in the art and analogous to the methods for creating synthetic recombinants of poxviruses such as the vaccinia virus and avipox virus described in U.S. Pat. Nos. 4,769,330, 4,722,848, 4,603,112, 5,110,587, and 5,174,993, the disclosures of which are incorporated herein by reference.

First, the DNA gene sequence to be inserted into the virus, particularly an open reading frame from a non-pox source, is placed into an E. coli plasmid construct into which DNA homologous to a section of DNA of the poxvirus has been inserted. Separately, the DNA gene sequence to be inserted is ligated to a promoter. The promoter-gene linkage is positioned in the plasmid construct so that the promoter-gene linkage is flanked on both ends by DNA homologous to a DNA sequence flanking a region of pox DNA containing a nonessential locus. The resulting plasmid construct is then amplified by growth within E. coli bacteria (Clewell, 1972) and isolated (Clewell et al., 1969; Maniatis et al., 1982).

Second, the isolated plasmid containing the DNA gene sequence to be inserted is transfected into a cell culture, e.g. chick embryo fibroblasts, along with the poxvirus. Recombination between homologous pox DNA in the plasmid and the viral genome respectively gives a poxvirus modified by the presence, in a nonessential region of its genome, of foreign DNA sequences. The term "foreign" DNA designates exogenous DNA, particularly DNA from a non-pox source, that codes for gene products not ordinarily produced by the genome into which the exogenous DNA is placed.

Genetic recombination is in general the exchange of homologous sections of DNA between two strands of DNA. In certain viruses RNA may replace DNA. Homologous sections of nucleic acid are sections of nucleic acid (DNA or RNA) which have the same sequence of nucleotide bases.

Genetic recombination may take place naturally during the replication or manufacture of new viral genomes within the infected host cell. Thus, genetic recombination between viral genes may occur during the viral replication cycle that takes place in a host cell which is co-infected with two or more different viruses or other genetic constructs. A section of DNA from a first genome is used interchangeably in constructing the section of the genome of a second co-infecting virus in which the DNA is homologous with that of the first viral genome.

However, recombination can also take place between sections of DNA in different genomes that are not perfectly homologous. If one such section is from a first genome homologous with a section of another genome except for the presence within the first section of, for example, a genetic marker or a gene coding for an antigenic determinant inserted into a portion of the homologous DNA, recombination can still take place and the products of that recombination are then detectable by the presence of that genetic marker or gene in the recombinant viral genome. Additional strategies have recently been reported for generating recombinant vaccinia virus.

Successful expression of the inserted DNA genetic sequence by the modified infectious virus requires two conditions. First, the insertion must be into a nonessential region of the virus in order that the modified virus remain viable. The second condition for expression of inserted DNA is the presence of a promoter in the proper relationship to the inserted DNA. The promoter must be placed so that it is located upstream from the DNA sequence to be expressed.

Vaccinia virus has been used successfully to immunize against smallpox, culminating in the worldwide eradication of smallpox in 1980. In the course of its history, many strains of vaccinia have arisen. These different strains demonstrate varying immunogenicity and are implicated to varying degrees with potential complications, the most serious of which are post-vaccinial encephalitis and generalized vaccinia (Behbehani, 1983).

With the eradication of smallpox, a new role for vaccinia became important, that of a genetically engineered vector for the expression of foreign genes. Genes encoding a vast number of heterologous antigens have been expressed in vaccinia, often resulting in protective immunity against challenge by the corresponding pathogen (reviewed in Tartaglia et al., 1990a).

The genetic background of the vaccinia vector has been shown to affect the protective efficacy of the expressed foreign immunogen. For example, expression of Epstein Barr Virus (EBV) gp340 in the Wyeth vaccine strain of vaccinia virus did not protect cottontop tamarins against EBV virus induced lymphoma, while expression of the same gene in the WR laboratory strain of vaccinia virus was protective (Morgan et al., 1988).

A fine balance between the efficacy and the safety of a vaccinia virus-based recombinant vaccine candidate is extremely important. The recombinant virus must present the immunogen(s) in a manner that elicits a protective immune response in the vaccinated animal but lacks any significant pathogenic properties. Therefore attenuation of the vector strain would be a highly desirable advance over the current state of technology.

A number of vaccinia genes have been identified which are non-essential for growth of the virus in tissue culture and whose deletion or inactivation reduces virulence in a variety of animal systems.

The gene encoding the vaccinia virus thymidine kinase (TK) has been mapped (Hruby et al., 1982) and sequenced (Hruby et al., 1983; Weir et al., 1983). Inactivation or complete deletion of the thymidine kinase gene does not prevent growth of vaccinia virus in a wide variety of cells in tissue culture. TK⁻ vaccinia virus is also capable of replication in vivo at the site of inoculation in a variety of hosts by a variety of routes.

It has been shown for herpes simplex virus type 2 that intravaginal inoculation of guinea pigs with TK⁻ virus resulted in significantly lower virus titers in the spinal cord than did inoculation with TK⁺ virus (Stanberry et al., 1985). It has been demonstrated that herpesvirus encoded TK activity in vitro was not important for virus growth in actively metabolizing cells, but was required for virus growth in quiescent cells (Jamieson et al., 1974).

Attenuation of TK⁻ vaccinia has been shown in mice inoculated by the intracerebral and intraperitoneal routes (Buller et al., 1985). Attenuation was observed both for the WR neurovirulent laboratory strain and for the Wyeth vaccine strain. In mice inoculated by the intradermal route, TK⁻ recombinant vaccinia generated equivalent anti-vaccinia neutralizing antibodies as compared with the parental TK⁺ vaccinia virus, indicating that in this test system the loss of TK function does not significantly decrease immunogenicity of the vaccinia virus vector. Following intranasal inoculation of mice with TK⁻ and TK⁺ recombinant vaccinia virus (WR strain), significantly less dissemination of virus to other locations, including the brain, has been found (Taylor et al., 1991a).

Another enzyme involved with nucleotide metabolism is ribonucleotide reductase. Loss of virally encoded ribonucleotide reductase activity in herpes simplex virus (HSV) by deletion of the gene encoding the large subunit was shown to have no effect on viral growth and DNA synthesis in dividing cells in vitro, but severely compromised the ability of the virus to grow on serum starved cells (Goldstein et al., 1988). Using a mouse model for acute HSV infection of the eye and reactivatable latent infection in the trigeminal ganglia, reduced virulence was demonstrated for HSV deleted of the large subunit of ribonucleotide reductase, compared to the virulence exhibited by wild type HSV (Jacobson et al., 1989).

Both the small (Slabaugh et al., 1988) and large (Schmidtt et al., 1988) subunits of ribonucleotide reductase have been identified in vaccinia virus. Insertional inactivation of the large subunit of ribonucleotide reductase in the WR strain of vaccinia virus leads to attenuation of the virus as measured by intracranial inoculation of mice (Child et al., 1990).

The vaccinia virus hemagglutinin gene (HA) has been mapped and sequenced (Shida, 1986). The HA gene of vaccinia virus is nonessential for growth in tissue culture (Ichihashi et al., 1971). Inactivation of the HA gene of vaccinia virus results in reduced neurovirulence in rabbits inoculated by the intracranial route and smaller lesions in rabbits at the site of intradermal inoculation (Shida et al., 1988). The HA locus was used for the insertion of foreign genes in the WR strain (Shida et al., 1987), derivatives of the Lister strain (Shida et al., 1988) and the Copenhagen strain (Guo et al., 1989) of vaccinia virus. Recombinant HA⁻ vaccinia virus expressing foreign genes have been shown to be immunogenic (Guo et al., 1989; Itamura et al., 1990; Shida et al., 1988; Shida et al., 1987) and protective against challenge by the relevant pathogen (Guo et al., 1989; Shida et al., 1987).

Cowpox virus (Brighton red strain) produces red (hemorrhagic) pocks on the chorioallantoic membrane of chicken eggs. Spontaneous deletions within the cowpox genome generate mutants which produce white pocks (Pickup et al., 1984). The hemorrhagic function (u) maps to a 38 kDa protein encoded by an early gene (Pickup et al., 1986). This gene, which has homology to serine protease inhibitors, has been shown to inhibit the host inflammatory response to cowpox virus (Palumbo et al., 1989) and is an inhibitor of blood coagulation.

The u gene is present in WR strain of vaccinia virus (Kotwal et al., 1989b). Mice inoculated with a WR vaccinia virus recombinant in which the u region has been inactivated by insertion of a foreign gene produce higher antibody levels to the foreign gene product compared to mice inoculated with a similar recombinant vaccinia virus in which the u gene is intact (Zhou et al., 1990). The u region is present in a defective nonfunctional form in Copenhagen strain of vaccinia virus (open reading frames B13 and B14 by the terminology reported in Goebel et al., 1990a,b).

Cowpox virus is localized in infected cells in cytoplasmic A type inclusion bodies (ATI) (Kato et al., 1959). The function of ATI is thought to be the protection of cowpox virus virions during dissemination from animal to animal (Bergoin et al., 1971). The ATI region of the cowpox genome encodes a 160 kDa protein which forms the matrix of the ATI bodies (Funahashi et al., 1988; Patel et al., 1987). Vaccinia virus, though containing a homologous region in its genome, generally does not produce ATI. In WR strain of vaccinia, the ATI region of the genome is translated as a 94 kDa protein (Patel et al., 1988). In Copenhagen strain of vaccinia virus, most of the DNA sequences corresponding to the ATI region are deleted, with the remaining 3' end of the region fused with sequences upstream from the ATI region to form open reading frame (ORF) A26L (Goebel et al., 1990a,b).

A variety of spontaneous (Altenburger et al., 1989; Drillien et al., 1981; Lai et al., 1989; Moss et al., 1981; Paez et al., 1985; Panicali et al., 1981) and engineered (Perkus et al., 1991; Perkus et al., 1989; Perkus et al., 1986) deletions have been reported near the left end of the vaccinia virus genome. A WR strain of vaccinia virus with a 10 kb spontaneous deletion (Moss et al., 1981; Panicali et al., 1981) was shown to be attenuated by intracranial inoculation in mice (Buller et al., 1985). This deletion was later shown to include 17 potential ORFs (Kotwal et al., 1988b). Specific genes within the deleted region include the virokine N1L and a 35 kDa protein (C3L, by the terminology reported in Goebel et al., 1990a,b). Insertional inactivation of NIL reduces virulence by intracranial inoculation for both normal and nude mice (Kotwal et al., 1989a). The 35 kDa protein is secreted like N1L into the medium of vaccinia virus infected cells. The protein contains homology to the family of complement control proteins, particularly the complement 4B binding protein (C4bp) (Kotwal et al., 1988a). Like the cellular C4bp, the vaccinia 35 kDa protein binds the fourth component of complement and inhibits the classical complement cascade (Kotwal et al., 1990). Thus the vaccinia 35 kDa protein appears to be involved in aiding the virus in evading host defense mechanisms.

The left end of the vaccinia genome includes two genes which have been identified as host range genes, K1L (Gillard et al., 1986) and C7L (Perkus et al., 1990). Deletion of both of these genes reduces the ability of vaccinia virus to grow on a variety of human cell lines (Perkus et al., 1990).

Two additional vaccine vector systems involve the use of naturally host-restricted poxviruses, avipox viruses. Both fowlpoxvirus (FPV) and canarypoxvirus (CPV) have been engineered to express foreign gene products. Fowlpox virus (FPV) is the prototypic virus of the Avipox genus of the Poxvirus family. The virus causes an economically important disease of poultry which has been well controlled since the 1920's by the use of live attenuated vaccines. Replication of the avipox viruses is limited to avian species (Matthews, 1982) and there are no reports in the literature of avipoxvirus causing a productive infection in any non-avian species including man. This host restriction provides an inherent safety barrier to transmission of the virus to other species and makes use of avipoxvirus based vaccine vectors in veterinary and human applications an attractive proposition.

FPV has been used advantageously as a vector expressing antigens from poultry pathogens. The hemagglutinin protein of a virulent avian influenza virus was expressed in an FPV recombinant (Taylor et al., 1988a). After inoculation of the recombinant into chickens and turkeys, an immune response was induced which was protective against either a homologous or a heterologous virulent influenza virus challenge (Taylor et al., 1988a). FPV recombinants expressing the surface glycoproteins of Newcastle Disease Virus have also been developed (Taylor et al., 1990; Edbauer et al., 1990).

Despite the host-restriction for replication of FPV and CPV to avian systems, recombinants derived from these viruses were found to express extrinsic proteins in cells of nonavian origin. Further, such recombinant viruses were shown to elicit immunological responses directed towards the foreign gene product and where appropriate were shown to afford protection from challenge against the corresponding pathogen (Tartaglia et al., 1993a,b; Taylor et al., 1992; 1991b; 1988b).

Human cytomegalovirus (HCMV) is a member of the betaherpesviridae subfamily (family Herpesviridae). HCMV is ubiquitous in humans, with usually mild or inapparent acute infection followed by persistence or latency. However, HCMV is a significant cause of morbidity and mortality in infants infected in-utero (Stagno et al., 1983). HCMV is the most common infectious complication of organ transplantation (Glenn et al., 1981) and in immunocompromised hosts (Weller et al., 1971). In AIDS patients, CMV retinitis is the leading cause of blindness (Roarty et al., 1993; Gallant et al., 1992; Gross et al., 1990) A potential role of HCMV in coronary restinosis has recently been described (Speir et al., 1994). The live attenuated Towne strain of HCMV has been shown to protect seronegative renal transplant recipients from severe clinical symptoms of HCMV infection (Plotkin et al., 1976, 1984 and 1989) and to protect initially seronegative healthy individuals from infection and clinical symptoms after subcutaneous challenge with a wild-type strain of HCMV (Plotkin et al., 1989). Concerns remain about the use of a live HCMV vaccine because of the latency reactivation phenomenon characteristic of herpesvirus infections in humans and because of the capability of certain strains of HCMV to transform cells malignantly in vitro (Albrecht and Rapp, 1973; Galloway et al., 1986). For these reasons, a recombinant subunit CMV vaccine may be more acceptable for human immunization.

The role of individual HCMV proteins in protective immunity is unclear. Three immunologically distinct families of glycoproteins associated with the HCMV envelope have been described (Gretch et al., 1988b); gCI (gp55 and gp93-130); gCII (gp47-52); and gCIII (gp85-p145). Neutralization of HCMV has been demonstrated in vitro with antibodies specific for each of these glycoprotein families (Pachl et al., 1989; Rasmussen et al., 1988; Kari et al., 1986).

The gene coding for gCI is homologous to HSV I gB (Cranage et al., 1986). HCMVgB is synthesized as a glycosylated uncleaved precursor of apparent molecular weight 130-140 kDa which is processed by cellular proteinase into N-terminal 90-110 kDa and C-terminal 55-58 kDa products which remain associated in a disulfide linked complex (Britt and Auger, 1986; Britt and Vugler, 1989; Reis et al., 1993). Monoclonal antibodies capable of neutralizing HCMV have been obtained from mice immunized with lysates of HCMV infected cells or HCMV virions, these monoclonals were predominantly reactive with the C-terminal 55-58 kDa fragment (Britt, 1984; Kari et al., 1986; Pereira et al., 1984; Rasmussen et al., 1988). However, immunization with biochemically purified gP93 resulted in the development of gp93-specific neutralizing mAbs (Kari et al., 1990).

HCMV-gB may serve to elicit protective immunity in humans: immunization with the purified gB protein induces neutralizing antibody (Gonczol et al., 1990) and human antigB monoclonal antibodies neutralize the virus (Masuho et al., 1987). Following natural infection neutralizing antibody specific for HCMV-gB is observed. When gB specific antibody is absorbed from human sera, HCMV neutralizing antibody titer is reduced significantly (50-88%, Gonczol et al., 1991; 0-98% median 48%, Marshall et al., 1992). There is also evidence for activation of helper T cells by the gB protein in naturally seropositive humans (Liu et al., 1991) and gB specific CTL has been detected in humans in some studies (Borysiewicz et al., 1988; Liu et al., 1991; Riddell, et al., 1991).

The gCII glycoproteins are encoded by a gene or genes in the US6 gene family (US6 through US11, Gretch et al., 1988a). These glycoproteins are recognized by human anti-HCMV antibody in sera from convalescent adults. However, sera from congenitally infected infants with persistent infection failed to react with gCII glycoproteins (Kari and Gehrz, 1990), suggesting that gCII may be important to human protective immune responses to HCMV.

The gP86 component of the gCIII complex is encoded by a gene that is homologous to HSV-I gH (Cranage et al., 1988; Pachl et al., 1989). The HCMV gH protein is capable of inducing a neutralizing immune response in humans (10% of HCMV infected individuals have a detectable level of circulating gH specific antibody (Rasmussen et al., 1991) as well as in laboratory animals (Baboonian et al., 1989; Cranage et al., 1988; Ehrlich et al., 1988; Rasmussen et al., 1984). Murine gH-specific monoclonal antibodies neutralize virus infectivity in a complement-independent manner (Baboonian et al., 1989; Cranage et al., 1988; Rasmussen et al., 1984) and inhibit viral spread (Pachl et al., 1989) suggesting that gH may be responsible for virus attachment, penetration and or spread.

Although gH is found on the surface of HCMV infected cells (Cranage et al., 1988), when expressed by a variety of recombinant systems it is restricted to the endoplasmic reticulum (Spaete et al., 1991). Coexpression of the HCMV UL115 gene product (glycoprotein gL) results in the formation of a stable complex of these two proteins and the transport of gH to the cell surface (Spaete et al., 1993; Kaye et al., 1992).

HCMV synthesizes a number of matrix tegument phosphoproteins. The pp150 phosphoprotein is highly immunogenic apparently more so than any other of the HCMV structural proteins (Jahn et al., 1987). A second matrix phosphoprotein, pp65, elicits a variable humoral response in humans (Jahn et al., 1987; Plachter et al., 1990). This protein can stimulate lymphoproliferation, IL-2 and interferon production, B-cell stimulation of antibody and natural killer cell activity (Forman et al., 1985). It also serves as a target antigen for HCMV-specific, HLA-restricted cytotoxic T cells (CTLs) (Pande et al., 1991; Gilbert et al., 1993).

Additional structural proteins may be required for eliciting a protective immune response to HCMV. The major capsid protein (UL86) is known to induce specific antibodies during natural infection and has been considered as the CMV-group common antigen (Spaete et al., 1994). The tegument phosphoprotein, pp28 (UL99), is also known to elicit persistent antibody responses during a natural infection. Further, this protein has also been implicated as a CTL target immunogen (Charpentier et al., 1986). The immune response to the upper tegument phosphoprotein, pp71 (UL82), is not as well characterized as the other tegument phosphoproteins (pp28, pp65), but as a known tegument protein requires further attention.

In addition to these structural proteins, some nonstructural proteins may also be candidates for inclusion in a recombinant subunit vaccine. Immunization of mice with a recombinant vaccinia virus expressing murine cytomegalovirus (MCMV) pp89 (functional homolog of HCMV IE 1) induces CD8⁺ T-cell responses that mediate protective immunity from challenge with MCMV (Jonjic et al., 1988). The human CMV major immediate early protein (IE 1) has been shown to be a target for CTLs isolated from HCMV seropositive individuals (Borysiewicz et al., 1988). Since IE 1 is among the initial viral proteins expressed and is necessary for inducing the expression of other CMV genes and initiating the viral life cycle in latently infected cells (Blanton and Tevethia, 1981; Cameron and Preston, 1981; DeMarchi et al., 1980: McDonough and Spector, 1983; Wathen et al., 1981), CTL responses directed against IE 1 may be important for controlling and/or eliminating HCMV infection. Recently Gilbert et al., (1993) have suggested that HCMV has evolved a mechanism by which other viral encoded proteins selectively interfere with the presentation of IE-derived peptides in association with Class I major histocompatibility complex (MHC) molecules.

Some additional nonstructural proteins may also be candidates for inclusion in a recombinant subunit HCMV vaccine candidate. The immediate early protein, IE2 (UL122), and the regulatory protein UL69 are known to contain human T-helper epitopes (Beninga et al., 1995).

One approach to the development of a subunit HCMV vaccine is the use of live viral vectors to express relevant HCMV gene products.

It can thus be appreciated that provision of a CMV or an HCMV recombinant poxvirus, and of compositions and products therefrom particularly NYVAC or ALVAC based CMV or HCMV recombinants and compositions and products therefrom, especially such recombinants containing coding for any or all of HCMVgB, gH, gL, pp150, pp65 and IE1, including recombinants expressing altered or truncated versions of IE1 and/or gB and compositions and products therefrom would be a highly desirable advance over the current state of technology.

OBJECTS AND SUMMARY OF THE INVENTION

It is therefore an object of this invention to provide modified recombinant viruses, which viruses have enhanced safety, and to provide a method of making such recombinant viruses.

It is an additional object of this invention to provide a recombinant poxvirus antigenic vaccine or immunological composition having an increased level of safety compared to known recombinant poxvirus vaccines.

It is a further object of this invention to provide a modified vector for expressing a gene product in a host, wherein the vector is modified so that it has attenuated virulence in the host.

It is another object of this invention to provide a method for expressing a gene product in a cell cultured in vitro using a modified recombinant virus or modified vector having an increased level of safety.

These and other objects and advantages of the present invention will become more readily apparent after consideration of the following.

In one aspect, the present invention relates to a modified recombinant virus having inactivated virus-encoded genetic functions so that the recombinant virus has attenuated virulence and enhanced safety. The functions can be non-essential, or associated with virulence. The virus is advantageously a poxvirus, particularly a vaccinia virus or an avipox virus, such as fowlpox virus or canarypox virus. The modified recombinant virus can include, within a non-essential region of the virus genome, a heterologous DNA sequence which encodes an antigen or epitope derived from HCMV, such as any or all of HCMVgB, gH, gL, pp150, pp65, IE1, including altered or truncated versions of IE1, and/or gB.

In another aspect, the present invention relates to an antigenic, immunological or vaccine composition or a therapeutic composition for inducing an antigenic or immunological response in a host animal inoculated with the composition, said vaccine including a carrier and a modified recombinant virus having inactivated nonessential virus-encoded genetic functions so that the recombinant virus has attenuated virulence and enhanced safety. The virus used in the composition according to the present invention is advantageously a poxvirus, particularly a vaccinia virus or an avipox virus, such as fowlpox virus and canarypox virus. The modified recombinant virus can include, within a nonessential region of the virus genome, a heterologous DNA sequence which encodes an antigenic protein, e.g., derived from HCMV, such as any or all of HCMVgB, gH, gL, pp150, pp65, IE1, including altered or truncated versions of IE1, and/or gB.

In yet another aspect, the present invention relates to an immunogenic composition containing a modified recombinant virus having inactivated nonessential virus-encoded genetic functions so that the recombinant virus has attenuated virulence and enhanced safety. The modified recombinant virus includes, within a non-essential region of the virus genome, a heterologous DNA sequence which encodes an antigenic protein (e.g., derived from HCMV, such as any or all of HCMVgB, gH, gL, pp150, pp65, IE1, including altered or truncated versions of IE1, and/or gB) wherein the composition, when administered to a host, is capable of inducing an immunological response specific to the antigen.

In a further aspect, the present invention relates to a method for expressing a gene product in a cell in vitro by introducing into the cell a modified recombinant virus having attenuated virulence and enhanced safety. The modified recombinant virus can include, within a nonessential region of the virus genome, a heterologous DNA sequence which encodes an antigenic protein, e.g. derived from HCMV such as any or all of HCMVgB, gH, gL, pp150, pp65, IE1, including altered or truncated versions of IE1, and/or gB. The cells can then be reinfused directly into the individual or used to amplify specific reactivities for reinfusion (Ex vivo therapy).

In a further aspect, the present invention relates to a method for expressing a gene product in a cell cultured in vitro by introducing into the cell a modified recombinant virus having attenuated virulence and enhanced safety. The modified recombinant virus can include, within a nonessential region of the virus genome, a heterologous DNA sequence which encodes an antigenic protein, e.g., derived from HCMV such as any or all of HCMVgB, gH, gL, pp150, pp65, IE1, including altered or truncated versions of IE1, and/or gB. The product can then be administered to individuals or animals to stimulate an immune response. The antibodies raised can be useful in individuals for the prevention or treatment of HCMV and, the antibodies from individuals or animals or the isolated in vitro expression products can be used in diagnostic kits, assays or tests to determine the presence or absence in a sample such as sera of HCMV or antigens therefrom or antibodies thereto (and therefore the absence or presence of the virus or of the products, or of an immune response to the virus or antigens).

In a still further aspect, the present invention relates to a modified recombinant virus having nonessential virus-encoded genetic functions inactivated therein so that the virus has attenuated virulence, and wherein the modified recombinant virus further contains DNA from a heterologous source in a nonessential region of the virus genome. The DNA can code for HCMV such as any or all of HCMVgB, gH, gL, pp150, pp65, IE1, including altered or truncated versions of IE1, and/or gB. In particular, the genetic functions are inactivated by deleting an open reading frame encoding a virulence factor or by utilizing naturally host restricted viruses. The virus used according to the present invention is advantageously a poxvirus, particularly a vaccinia virus or an avipox virus, such as fowlpox virus or canarypox virus. Advantageously, the open reading frame is selected from the group consisting of J2R, B13R+B14R, A26L, A56R, C7L-K1L, and I4L (by the terminology reported in Goebel et al., 1990a,b); and, the combination thereof. In this respect, the open reading frame comprises a thymidine kinase gene, a hemorrhagic region, an A type inclusion body region, a hemagglutinin gene, a host range gene region or a large subunit, ribonucleotide reductase; or, the combination thereof. A suitable modified Copenhagen strain of vaccinia virus is identified as NYVAC (Tartaglia et al., 1992), or a vaccinia virus from which has been deleted J2R, B13R+B14R, A26L, A56R, C7L-K1l and I4L or a thymidine kinase gene, a hemorrhagic region, an A type inclusion body region, a hemagglutinin gene, a host range region, and a large subunit, ribonucleotide reductase (See also U.S. Pat. No. 5,364,773). Alternatively, a suitable poxvirus is an ALVAC or, a canarypox virus (Rentschler vaccine strain) which was attenuated, for instance, through more than 200 serial passages on chick embryo fibroblasts, a master seed therefrom was subjected to four successive plaque purifications under agar from which a plaque clone was amplified through five additional passages.

The invention in yet a further aspect relates to the product of expression of the inventive recombinant poxvirus and uses therefor, such as to form antigenic, immunological or vaccine compositions for treatment, prevention, diagnosis or testing; and, to DNA from the recombinant poxvirus which is useful in constructing DNA probes and PCR primers.

These and other embodiments are disclosed or are obvious from and encompassed by the following detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

The following detailed description, given by way of example, but not intended to limit the invention solely to the specific embodiments described, may best be understood in conjunction with the accompanying drawings, in which:

FIG. 1 schematically shows a method for the construction of plasmid pSD460 for deletion of thymidine kinase gene and generation of recombinant vaccinia virus vP410;

FIG. 2 schematically shows a method for the construction of plasmid pSD486 for deletion of hemorrhagic region and generation of recombinant vaccinia virus vP553;

FIG. 3 schematically shows a method for the construction of plasmid pMP494Δ for deletion of ATI region and generation of recombinant vaccinia virus vP618;

FIG. 4 schematically shows a method for the construction of plasmid pSD467 for deletion of hemagglutinin gene and generation of recombinant vaccinia virus vP723;

FIG. 5 schematically shows a method for the construction of plasmid pMPCK1Δ for deletion of gene cluster [C7L-K1L] and generation of recombinant vaccinia virus vP804;

FIG. 6 schematically shows a method for the construction of plasmid pSD548 for deletion of large subunit, ribonucleotide reductase and generation of recombinant vaccinia virus vP866 (NYVAC);

FIG. 7 schematically shows a method for the construction of plasmid pRW842 for insertion of rabies glycoprotein G gene into the TK deletion locus and generation of recombinant vaccinia virus vP879;

FIG. 8 shows the DNA sequence of a 3209 base pair fragment of canarypox DNA containing the C5 ORF (SEQ ID NO:27) (the C5 ORF initiates at position 1537 and terminates at position 1857);

FIGS. 9A and 9B schematically show a method for the construction of recombinant canarypox virus vCP65 (ALVACRG);

FIG. 10 shows schematically the ORFs deleted to generate NYVAC;

FIGS. 11A to 11D show graphs of rabies neutralizing antibody titers (RFFIT, IU/ml), booster effect of HDC and vCP65 (10⁵.5 TCID₅₀) in volunteers previously immunized with either the same or the alternate vaccine (vaccines given at days 0, 28 and 180, antibody titers measured at days 0, 7, 28, 35, 56, 173, 187 and 208);

FIG. 12 shows the DNA sequence of HCMVgB (Towne strain) (SEQ ID NO:37);

FIGS. 13A and B show the DNA sequence of the H6 promoted HCMVgB and NYVAC sequences flanking the TK locus (SEQ ID NO:38) (the 5'end of the H6 promoted CMVgB is at position 3447; the CMVgB coding sequence is from position 3324 through position 606);

FIGS. 14A to C show the DNA sequence of a 7351 base pair fragment of canarypox DNA containing the C3 ORF (SEQ ID NO:39) (the C3 ORF is initiated at position 1458 and terminates at position 2897);

FIGS. 15A to C show the DNA sequence of the H6 promoted HCMVgB and ALVAC sequences flanking the C3 locus (SEQ ID NO:40) (the 5' end of the H6 promoted CMVgB is at position 4425; the CMVgB coding sequence is from position 4301 through position 1581);

FIGS. 16A and B show the DNA sequence of the H6 promoted HCMVgB and NYVAC sequences flanking the ATI locus (SEQ ID NO:41) (the 5'end of the H6 promoted CMVgB is at position 3348; the CMVgB coding sequence is from position 3224 through position 504);

FIG. 17 shows the DNA sequence of HCMVgB (Towne strain) deleted of its transmembrane region (SEQ ID NO:42);

FIGS. 18A and B show the DNA sequence of the H6 promoted HCMVgB lacking its transmembrane region and NYVAC sequences flanking the ATI locus (SEQ ID NO:43) (the 5' end of the H6 promoted CMVgB is at position 3173; the CMVgB coding sequence is from position 3050 through position 504);

FIG. 19 shows the DNA sequence of HCMVgB (Towne strain) deleted of its transmembrane region and containing an altered cleavage site (SEQ ID NO:44);

FIGS. 20A and B show the DNA sequence of the H6 promoted HCMVgB lacking its transmembrane region and containing an altered cleavage site plus NYVAC sequences flanking the ATI locus (SEQ ID NO:45) (the 5' end of the H6 promoted CMVgB is at position 3173; the CMVgB coding sequence is from position 3050 through position 504);

FIG. 21 shows the DNA sequence of HCMVgH (Towne strain) (SEQ ID NO:46);

FIGS. 22A and B show the DNA sequence of the 42K promoted HCMVgH plus NYVAC sequences flanking the I4L locus (SEQ ID NO:47) (the 5' end of the 42K promoted CMVgH is at position 641; the CMVgH coding sequence is from position 708 through position 2933);

FIGS. 23A and B show the DNA sequence of the 42K promoted CMVgH and ALVAC sequences flanking the C5 locus (SEQ ID NO:48) (the 5' end of the 42K promoted CMVgH is at position 1664; the CMVgH coding sequence is from position 1730 through position 3955);

FIG. 24 shows the DNA sequence of the 42K promoted CMVgH and WR flanking sequences (SEQ ID NO:49) (the 5' end of the 42K promoted CMVgH is at position 2457; the CMVgH coding sequence is from position 2391 through 166);

FIG. 25 shows the DNA sequence of HCMV IE1 (AD169 strain) (SEQ ID NO:50);

FIG. 26 shows the DNA sequence of the H6 promoted CMVIE1 and WR flanking sequences (SEQ ID NO:51) (the 5' end of the H6 promoted CMVIE1 is at position 1796; the CMVIE1 coding sequence is from position 1673 through 201);

FIGS. 27A and B show the DNA sequence of the H6 promoted CMVIE1 and NYVAC sequences flanking the ATI locus (SEQ ID NO:52) (the 5' end of the H6 promoted CMVIE1 is at position 2030; the CMVIE1 coding sequence is from position 1906 through position 434); FIG. 28 shows the DNA sequence of HCMVIE1 (AD169 strain) lacking amino acids 292-319 (SEQ ID NO:53);

FIGS. 29A and B show the DNA sequence of the H6 promoted CMVIE1 lacking amino acids 292-319 and NYVAC sequences flanking the ATI locus (SEQ ID NO:54) (the 5' end of the H6 promoted CMVIE1 is at position 1940; the CMVIE1 coding sequence is from position 1816 through position 434);

FIG. 30 shows the DNA sequence of the Exon 4 segment of HCMVIE1 (AD169 strain) (SEQ ID NO:55);

FIG. 31 shows the DNA sequence of the H6 promoted CMVIE1 Exon 4 segment and NYVAC sequences flanking the I4L locus (SEQ ID NO:56) (the 5' end of the H6 promoted IE1 Exon 4 is at position 630; the CMVIE1 Exon 4 coding sequence is from position 754 through position 1971).

FIG. 32A and B show the DNA sequence of the H6 promoted CMVIE1 Exon 4 segment and ALVAC sequences flanking the C5 locus (SEQ ID NO:57) (the 5' end of the H6 promoted IE1 Exon 4 is at position 1647; the CMVIE1 Exon 4 coding sequence is from position 1771 through position 2988).

FIG. 33 shows the DNA sequence of HCMVIE1 (AD169 strain) lacking amino acids 2-32 (SEQ ID NO:58);

FIG. 34 shows the DNA sequence of the H6 promoted CMVIE1 lacking amino acids 2-32 and NYVAC sequences flanking the I4L locus (SEQ ID NO:59) (the 5' end of the H6 promoted IE1 lacking amino acids 2-32 is at position 630; the coding sequence for CMVIE1 lacking amino acids 2-32 is from position 754 through position 2133);

FIGS. 35A and B show the DNA sequence of the H6 promoted CMVIE1 lacking amino acids 2-32 and ALVAC sequences flanking the C5 locus (SEQ ID NO:60) (the 5' end of the H6 promoted IE1 lacking amino acids 2-32 is at position 1647; the CMVIE1 coding sequence for CMVIE1 lacking amino acids 2-32 is from position 1771 through position 3150);

FIG. 36 shows the DNA sequence of HCMV pp65 (Towne strain) (SEQ ID NO:61);

FIG. 37 shows the DNA sequence of the H6 promoted CMVpp65 and NYVAC sequences flanking the HA locus (SEQ ID NO:62) (the 5' end of the H6 promoted pp65 is at position 476; the CMVpp65 coding sequence is from position 600 through 2282);

FIGS. 38A and B show the DNA sequence of a 3706 base pair fragment of canarypox DNA containing the C6 ORF (SEQ ID NO:63) (the C6 ORF is initiated at position 377 and terminated at position 2254);

FIGS. 39A and B show the DNA sequence of the H6 promoted CMVpp65 and ALVAC sequences flanking the C6 locus (SEQ ID NO:64) (the 5' end of the H6 promoted pp65 is at position 496; the CMVpp65 coding sequence is from position 620 through 2302);

FIG. 40 shows the DNA sequence of the H6 promoted CMVpp65 and WR flanking sequences (SEQ ID NO:65) (the 5' end of the H6 promoted pp65 is at position 168; the CMVpp65 coding sequence is from position 292 through 1974);

FIG. 41 shows the DNA sequence of HCMVpp150 (Towne strain) (SEQ ID NO:66);

FIGS. 42A and B show the DNA sequence of the 42K promoted CMVpp150 and NYVAC sequences flanking the ATI locus (SEQ ID NO:67) (the 5' end of the 42K promoted pp150 is at position 3645; the CMVpp150 coding sequence is from position 3580 through 443);

FIGS. 43A and B show the DNA sequence of the 42K promoted CMVpp150 and ALVAC sequences flanking the C6 locus (SEQ ID NO:68) (the 5' end of the 42K promoted pp150 is at position 3714; the CMVpp150 coding sequence is from position 3649 through 512);

FIGS. 44A and B show the DNA sequence of the 42K promoted CMVpp150 gene and WR flanking sequences (SEQ ID NO:69) (the 5' end of the H6 promoted pp150 is at position 3377; the CMVpp150 coding sequence is from position 3312 through 175); FIGS. 45A and B show the DNA sequence of the 42K promoted HCMVgH and H6 promoted HCMVIE Exon 4 and NYVAC sequences flanking the I4L locus (SEQ ID NO:70) (the 5' end of the 42K promoted CMVgH is at position 2935; the CMVgH coding sequence is from position 2869 through 644; the 5' end of the H6 promoted CMVIE Exon 4 is at position 2946; the CMVIE Exon 4 coding sequence is from position 3070 through position 4287);

FIGS. 46A to C show the DNA sequence of the H6 promoted HCMV pp65 and 42K promoted HCMVpp150 and ALVAC sequences flanking the C6 locus (SEQ ID NO:71) (the 5' end of the H6 promoted CMVpp65 is at position 496; the CMVpp65 coding sequence is from position 620 through 2302; the 5' end of the 42K promoted CMVpp150 is at position 5554; the CMVpp150 coding sequence is from position 5489 through position 2352);

FIG. 47 shows the DNA sequence of HCMVgL (Towne strain) (SEQ ID NO:72);

FIGS. 48A and B show the DNA sequence of the H6 promoted HCMVgB and H6 promoted HCMVgL and NYVAC sequences flanking the TK locus (SEQ ID NO:73) (the 5' end of the H6 promoted CMVgB is at position 3447; the CMVgB coding sequence is from position 3324 through position 606; the 5' end of the H6 promoted CMVgL is at position 3500; the CMVgL coding sequence is from position 3624 through position 4460);

FIG. 49 shows the results of HCMV IE1 CTL stimulation by ALVAC-IE1 (vCP256) (percent cytotoxicity; white bars=WR, black bars=WRIE1, striped bars=nonautologous);

FIG. 50 shows the results of stimulation of HCMV pp65-CTLs by ALVAC-pp65 (vCP260) (human CTLs stimulated in vitro and assayed for HCMV pp65 CTLs using methodology similar to that used for FIG. 49; percent cytotoxity; white bars=WR, black bars=WR-pp65, striped bars=nonautologous);

FIG. 51 shows the results of stimulation of HCMV IE1 CTLs by ALVAC-IE1 (vCP256) (methodology similar to that used for FIG. 49, except that following 6 days incubation for restimulation, the responder mononuclear cells were incubated with immunomagnetic beads coupled to monoclonal anti-human CD3, CD4 or CD8; percent cytotoxicity; white bars=WR, black bars=WR-IE1, striped bars=HLA mismatch);

FIGS. 52A to D show expression of CMV gB by COPAK recombinants in Vero and HeLa cells (cell and medium fractions from infected cells radiolabeled with [S 35] methionine were immune precipitated with guinea pig anti-CMV gB; Vero medium (A), HeLa medium (B), Vero cell (C), and HeLa cell (D) fractions derived from infections by vP993 COPAK parent (lanes 1), vP1126 expressing the entire gB (lanes 2), vP1128 expressing gB without the transmembrane site (lanes 3), and vP1145 expressing the gB without transmembrane and with altered cleavage sites (lanes 4) are shown; far right lane contains molecular weight markers);

FIGS. 53A and B show vaccinia infection of Vero and HeLa cells detected by expression of vaccinia early protein E3L (cell fractions from infected cells radiolabeled with [35 S] methionine were immune precipitated with rabbit anti-p25 (E3L); Vero (A) and HeLa (B) cell fractions derived from infections by vP993 (lanes 1), vP1126 (lanes 2), vP1128 (lanes 3), and vP1145 (lanes 4) are shown; far right lane contains molecular weight markers);

FIG. 54 shows comparison of CMV gB production by Vero, HeLa and MRC-5 cells (SDS-PAGE and western blot analysis were performed on the medium from MRC-5 cells (lanes 1, 4), Vero cells (lanes 2, 5), or HeLa cells (lanes 3, 6) after infection with vP1145 (lanes 1, 2, 3) or vP993 (lanes 4, 5, 6); CMV gB was detected with monoclonal CH380; molecular weight markers are present in lane M);

FIG. 55 shows immunoprecipitation of CMV gB by a panel of monoclonal antibodies and guinea pig anti-gB (radiolabeled medium fractions from Vero cells infected with vP993 (lanes 1), vP1126 (lanes 2), vP1128 (lanes 3), and vP1145 (lanes 4) were immune precipitated with guinea pig anti-CMV gB or with monoclonals 13-127, 13-128, CH380, HCMV 34, or HCMV 37; far left lane contains molecular weight markers);

FIG. 56 shows western blot analysis of fractions and bed material from CMV gB immunoaffinity chromatography columns (column 19 fractions representing eluted gB (lane 5), flow through material (lane 6), and crude gB material applied to the column (lane 7) were analyzed by SDS-PAGE and western blot using monoclonal CH380; included in the assay was bed material from column 19 (lane 2) and column 11 (lane 3), as well as gB purified on column 7 (lane 4); molecular weight markers are present in lane 1);

FIG. 57 shows SDS-PAGE analysis of CMV gB eluted from an immunoaffinity chromatography column (fractions 8.16 through 8.22, eluted from column 8, were electrophoretically separated on a 10% gel under reducing conditions, and stained with silver);

FIG. 58 shows SDS-PAGE analysis of five batches of immunoaffinity purified CMV gB (samples of batches 1 through 5 (lanes 1-5) were electrophoretically separated on a 10% gel under reducing conditions and stained with Coomassie Blue; Lane M contains molecular weight markers);

FIGS. 59, 59A shows characterization of immunoaffinity purified CMV gB (batch 5, analyzed by SDS-PAGE, as shown in FIGS. 58A and B, was scanned with a densitometer, and bands were defined (lane 7, labels 1 through 8) with FIG. 59A showing a densitometer tracing through lane 7);

FIGS. 60A and B show immunoblot analysis of immunoaffinity purified CMV gB (purified HIV env (lanes 1), affinity purified CMV gB (lanes 2), crude CMV gB (lane (B3), or monoclonal CH380 (lane A3) were electrophoretically separated on a 10% gel, blotted onto nitrocellulose paper and probed for the presence of mouse IgG H and L chains or CMVgB using goat anti-mouse IgG (A) or monoclonal CH380 (B), respectively; molecular weight markers are present in lanes 4);

FIGS. 61A and B show immunoprecipitation/immunoblot analysis of affinity purified gB (Batch 1 immunoaffinity purified gB(1) or crude gB (B) was immunoprecipitated with monoclonals CH380 (lanes 1), 13-127 (lanes 2), 13-128 (lanes 3), HCMV 37 (lanes 4), or HCMV 34 (lanes 5); the immunoprecipitates were electrophoretically separated on a 10% gel under reducing conditions, blotted onto nitrocellulose and probed for the presence of gB, using guinea pig anti-CMB gB; far left lanes are molecular weight markers);

FIGS. 62A and B show immunoblot analysis of affinity purified CMV gB (Vero cells lysates (lanes A3, B2), CEF lysates (lane A2), vaccinia-infected Vero cells (lane B3), crude CMV gB (lanes 4), affinity purified CMV gB (lanes 5), or purified HIV env (lanes 6) were electrophoretically separated on a 10% gel under reducing conditions, blotted onto nitrocellulose, and probed for the presence of Vero cell proteins using rabbit anti-Vero cells (A), or vaccinia proteins using rabbit anti-vaccinia (B); molecular weight markers are present in lanes 1);

FIGS. 63A-C show the DNA sequence of the H6 promoted HCMVpp65 and 42K promoted HCMVpp150 and ALVAC sequences flanking the C6 locus (SEQ ID NO: 188) (The 5' end of the H6 promoted CMVpp65 is at position 496. The CMVpp65 coding sequence is from position 620 through 2302. The 5' end of the 42K promoted CMVpp150 is at position 2341. The CMVpp150 coding sequence is from position 2406 through 5543);.

FIGS. 64A and B show the DNA sequence of a 5798bp fragment of canarypox DNA containing the C₇ ORF (tk) (SEQ ID NO: 189) (The C₇ ORF is initiated at position 4412 and terminated at position 4951);

FIG. 65A and B show the DNA sequence of the H6 promoted HCMVgL gene and ALVAC sequences flanking the C₇ locus (The 5' end of the H6 promoted CMVgL gene is at position 2136. The CMVgL coding sequence is from position 2260 through 3093);

FIGS. 66A and B show the DNA sequence of the H6 promoted HCMVgL gene and H6 promoted HCMV IE1-exon4 gene and ALVAC sequences flanking the C₇ locus (SEQ ID NO: 190) (The 5' end of the H6 promoted CMVgL gene is at position 3476. The CMVgL coding region is from position 3600 through 4433. The 5' end of the H6 promoted IE1-exon4 is at position 3469. The CMV IE1-exon4 coding region is from position 3345 through 2128);

FIG. 67 shows the DNA sequence of HCMVgH (SEQ ID NO: 191)(Towne strain) deleted of its transmembrane region and cytoplasmic tail; and

FIGS. 68A and B show the DNA sequence of the H6 promoted HCMVgL gene and 42K promoted truncated HCMVgH gene and NYVAC sequences flanking the ATI locus (SEQ ID NO: 191) (The 5' end of the H6 promoted CMVgL gene is at position 2669. The CMVgL coding region is from position 2793 through 3626. The 5' end of the 42K promoted truncated CMVgH gene is at position 2650. The truncated CMVgH coding sequence is from position 2584 through 434).

DETAILED DESCRIPTION OF THE INVENTION

To develop a new vaccinia vaccine strain, NYVAC (vP866), the Copenhagen vaccine strain of vaccinia virus was modified by the deletion of six nonessential regions of the genome encoding known or potential virulence factors. The sequential deletions are detailed below (See U.S. Pat. No. 5,364,773). All designations of vaccinia restriction fragments, open reading frames and nucleotide positions are based on the terminology reported in Goebel et al., 1990a,b.

The deletion loci were also engineered as recipient loci for the insertion of foreign genes.

The regions deleted in NYVAC are listed below. Also listed are the abbreviations and open reading frame designations for the deleted regions (Goebel et al., 1990a,b) and the designation of the vaccinia recombinant (vP) containing all deletions through the deletion specified:

(1) thymidine kinase gene (TK; J2R) vP410;

(2) hemorrhagic region (u; B13R+B14R) vP553;

(3) A type inclusion body region (ATI; A26L) vP618;

(4) hemagglutinin gene (HA; A56R) vP723;

(5) host range gene region (C7L-K1L) vP804; and

(6) large subunit, ribonucleotide reductase (I4L) vP866 (NYVAC).

NYVAC is a genetically engineered vaccinia virus strain that was generated by the specific deletion of eighteen open reading frames encoding gene products some of which associated with virulence and host range (Tartaglia et al., 1992; Goebel et al., 1990a,b). The deletion of host range genes diminishes the ability of the virus to replicate in tissue culture cell derived from certain species such as swine and humans (Tartaglia et al., 1992; Perkus et al., 1990). In addition to reduced replication competency, NYVAC was shown to be highly attenuated by a number of criteria including (a) lack of induration or ulceration on rabbit skin, (b) rapid clearance from the site of inoculation, (c) high avirulence by intracranial inoculation into newborn mice when compared with other vaccinia strains including WYETH, and (d) failure to cause death, secondary lesions or disseminated infection when inoculated intraperitoneally in immunocompromised animals (Tartaglia et al., 1992). In spite of the highly attenuated characteristics of NYVAC, NYVAC based recombinants were effective in protecting mice from rabies challenge (Tartaglia et al., 1992), swine from challenge with Japanese encephalitis virus and pseudorabies virus challenge (Brockmeier et al., 1993; Konishi et al., 1992) and horses from equine influenza virus challenge (Taylor et al., 1993).

NYVAC is also highly attenuated by a number of criteria including i) decreased virulence after intracerebral inoculation in newborn mice, ii) inocuity in genetically (nu⁺ /nu⁺) or chemically (cyclophosphamide) immunocompromised mice, iii) failure to cause disseminated infection in immunocompromised mice, iv) lack of significant induration and ulceration on rabbit skin, v) rapid clearance from the site of inoculation, and vi) greatly reduced replication competency on a number of tissue culture cell lines including those of human origin. Nevertheless, NYVAC based vectors induce excellent responses to extrinsic immunogens and provided protective immunity.

Avipoxvirus-based recombinants as live vectors provide an additional approach to develop recombinant subunit vaccines. These viruses are naturally restricted by their ability to replicate only in avian species. TROVAC refers to an attenuated fowlpox that was a plaque-cloned isolate derived from the FP-1 vaccine strain of fowlpoxvirus which is licensed for vaccination of 1 day old chicks.

ALVAC is an attenuated canarypox virus-based vector that was a plaque-cloned derivative of the licensed canarypox vaccine, Kanapox (Tartaglia et al., 1992). ALVAC has some general properties which are the same as some general properties of Kanapox. ALVAC-based recombinant viruses expressing extrinsic immunogens have also been demonstrated efficacious as vaccine vectors (Tartaglia et al., 1993 a,b). For instance, mice immunized with an ALVAC recombinant expressing the rabies virus glycoprotein were protected from lethal challenge with rabies virus (Tartaglia et al., 1992) demonstrating the potential for ALVAC as a vaccine vector. ALVAC-based recombinants have also proven efficacious in dogs challenged with canine distemper virus (Taylor et al., 1992) and rabies virus (Perkus et al., 1994), in cats challenged with feline leukemia virus (Tartaglia et al., 1993b), and in horses challenged with equine influenza virus (Taylor et al., 1993).

This avipox vector is restricted to avian species for productive replication. On human cell cultures, canarypox virus replication is aborted early in the viral replication cycle prior to viral DNA synthesis. Nevertheless, when engineered to express extrinsic immunogens, authentic expression and processing is observed in vitro in mammalian cells and inoculation into numerous mammalian species induces antibody and cellular immune responses to the extrinsic immunogen and provides protection against challenge with the cognate pathogen (Taylor et al., 1992; Taylor et al., 1991b). Recent Phase I clinical trials in both Europe and the United States of a canarypox/rabies glycoprotein recombinant (ALVAC-RG; vCP65) demonstrated that the experimental vaccine was well tolerated and induced protective levels of rabiesvirus neutralizing antibody titers (Cadoz et al., 1992; Fries et al., 1992). Indeed, reactogenicity in volunteers following administration of ALVAC-RG was minimal; and following two administrations of ALVAC-RG at a dose of 10⁵.5 TCID₅₀, all vaccinees developed rabies neutralizing antibody. Additionally, peripheral blood mononuclear cells (PBMCs) derived from the ALVAC-RG vaccinates demonstrated significant levels of lymphocyte proliferation when stimulated with purified rabies virus (Fries et al., 1992).

An ALVAC recombinant expressing the HIV envelope glycoprotein gp160 (ALVAC-HIV; vCP125) has been tested in phase I human clinical trial in a prime/boost protocol with recombinant gp160 (Pialoux et al., 1995). Reactogenicity in volunteers following administration of ALVAC-HIV was minimal and this vaccine candidate primed both HIV-I envelope-specific humoral and cell-mediated immune responses.

Recent studies have indicated that a prime/boost protocol, whereby immunization with a poxvirus recombinant expressing a foreign gene product is followed by a boost using a purified subunit preparation form of that gene product, elicits an enhanced immune response relative to the response elicited with either product alone. Human volunteers immunized with a vaccinia recombinant expressing the HIV-1 envelope glycoprotein and boosted with purified HIV-1 envelope glycoprotein subunit preparation exhibit higher HIV-1 neutralizing antibody titers than individuals immunized with just the vaccinia recombinant or purified envelope glycoprotein alone (Graham et al., 1993; Cooney et al., 1993). Humans immunized with two injections of an ALVAC-HIV-1 env recombinant (vCP125) failed to develop HIV specific antibodies. Boosting with purified rgp160 from a vaccinia virus recombinant resulted in detectable HIV-1 neutralizing antibodies. Furthermore, specific lymphocyte T cell proliferation to rgp160 was clearly increased by the boost with rgp160. Envelope specific cytotoxic lymphocyte activity was also detected with this vaccination regimen (Pialoux et al., 1995). Macaques immunized with a vaccinia recombinant expressing the simian immunodeficiency virus (SIV) envelope glycoprotein and boosted with SIV envelope glycoprotein from a baculovirus recombinant are protected against a SIV challenge (Hu et al., 1991; 1992). In the same fashion, purified HCMvgB protein can be used in prime/boost protocols with NYVAC or ALVAC-gB recombinants.

NYVAC, ALVAC and TROVAC have also been recognized as unique among all poxviruses in that the National Institutes of Health ("NIH")(U.S. Public Health Service), Recombinant DNA Advisory Committee, which issues guidelines for the physical containment of genetic material such as viruses and vectors, i.e., guidelines for safety procedures for the use of such viruses and vectors which are based upon the pathogenicity of the particular virus or vector, granted a reduction in physical containment level: from BSL2 to BSL1. No other poxvirus has a BSL1 physical containment level. Even the Copenhagen strain of vaccinia virus--the common smallpox vaccine--has a higher physical containment level; namely, BSL2. Accordingly, the art has recognized that NYVAC, ALVAC and TROVAC have a lower pathogenicity than any other poxvirus.

ALVAC, TROVAC, and NYVAC were deposited under the terms of the Budapest Treaty with the American Type Culture Collection (ATCC), 12301 Parklawn Drive, Rockville, Md., 20852, USA:

NYVAC under ATCC accession number VR-2559 on Mar. 6, 1997;

TROVAC under ATCC accession number VR-2553 on Feb. 6, 1997

And, ALVAC under ATCC accession number VR-2547 on Nov. 14, 1996.

CMV is a frequent cause of morbidity and mortality in AIDS patients, bone marrow transplant recipients, and patients undergoing immunosuppressive therapies for neoplastic diseases. There is no effective, well-tolerated, pharmaceutical therapy for CMV infection. One approach might be the ex vivo stimulation of donor CMV-specific CTLs for the treatment and control of the often fatal pneumonia caused by CMV infection in the bone marrow transplant recipient. In fact, the treatment and control of CMV infection in man by adoptive transfer of CMV CTL clones has been successfully demonstrated (Riddell et al., 1992). However, in this instance, CMV was used to stimulate and maintain the CMV-specific CTL clones used in this therapeutic protocol. The use of CMV for the purpose of ex vivo stimulation of CTL clones has its drawbacks, the most obvious being the possibility of introducing additional CMV into an immunosuppressed patient. The availability of immunotherapeutic agents that provide a safe and acceptable means for stimulating antigen-specific cellular immune effector activities seems to be a major shortcoming in the field of adoptive immunotherapy. Protein subunits, although potentially safe, are notoriously poor at stimulating CTLS. Peptides, generally considered safe yet effective at stimulating a CTL response, are highly restrictive in their abilities to stimulate CTL responses. Peptides are typically capable of inducing a CTL response to only one CTL epitope of many possible CTL epitopes contained within a single protein. Furthermore, peptides typically stimulate CTL responses from only a restricted portion of the population, being restricted to only those individuals expressing a particular allele of the human major histocompatibility complex (MHC). Recombinant virus vectors are considered excellent inducers of CTL reactivities since they are capable of expressing the entire antigen, thus not restricted to a single epitope for a single segment of the population. However, most of these virus vectors, such as adenovirus, are capable of replication and are not considered safe for use in this type of protocol. Since ALVAC recombinants do not replicate in mammalian cells, yet are capable of stimulating antigen-specific CTL responses, as demonstrated by data contained within this application, ALVAC recombinants represent a uniquely safe and effective method for the ex vivo stimulation of virus-specific CTL clones for utilization in immunotherapeutic applications.

This invention pertains to NYVAC, ALVAC and vaccinia (WR strain) recombinants containing the HCMV genes encoding for gB, gH, gL, pp150, pp65 and IE 1, including truncated versions thereof, which are further described in the Examples below.

Clearly based on the attenuation profiles of the NYVAC, ALVAC, and TROVAC vectors and their demonstrated ability to elicit both humoral and cellular immunological responses to extrinsic immunogens (Tartaglia et al., 1993a,b; Taylor et al., 1992; Konishi et al., 1992) such recombinant viruses offer a distinct advantage over previously described vaccinia-based recombinant viruses.

The administration procedure for recombinant virus or expression product thereof, compositions of the invention such as immunological, antigenic or vaccine compositions or therapeutic compositions can be via a parenteral route (intradermal, intramuscular or subcutaneous). Such an administration enables a systemic immune response.

More generally, the inventive antigenic, immunological or vaccine compositions or therapeutic compositions (compositions containing the poxvirus recombinants of the invention) can be prepared in accordance with standard techniques well known to those skilled in the pharmaceutical art. Such compositions can be administered in dosages and by techniques well known to those skilled in the medical arts taking into consideration such factors as the age, sex, weight, and condition of the particular patient, and the route of administration. The compositions can be administered alone, or can be co-administered or sequentially administered with compositions of the invention or with other immunological, antigenic or vaccine or therapeutic compositions in seropositive individuals. The compositions can be administered alone, or can be co-administered or sequentially administered with compositions of the invention or with other antigenic, immunological, vaccine or therapeutic compositions in seronegative individuals. Such other compositions can include purified antigens from HCMV or from the expression of such antigens by a recombinant poxvirus or other vector system or, such other compositions can include a recombinant poxvirus which expresses other HCMV antigens or biological response modifiers again taking into consideration such factors as the age, sex, weight, and condition of the particular patient, and, the route of administration.

Examples of compositions of the invention include liquid preparations for orifice, e.g., oral, nasal, anal, vaginal, etc., administration such as suspensions, syrups or elixirs; and, preparations for parenteral, subcutaneous, intradermal, intramuscular or intravenous administration (e.g., injectable administration) such as sterile suspensions or emulsions. In such compositions the recombinant poxvirus may be in admixture with a suitable carrier, diluent, or excipient such as sterile water, physiological saline, glucose or the like.

Further, the products of expression of the inventive recombinant poxviruses can be used directly to stimulate an immune response in either seronegative or seropositive individuals or in animals. Thus, the expression products can be used in compositions of the invention instead or in addition to the inventive recombinant poxvirus in the aforementioned compositions.

Additionally, the inventive recombinant poxvirus and the expression products therefrom stimulate an immune or antibody response in humans and animals and therefore those products are antigens. From those antibodies or antigens, by techniques well-known in the art, monoclonal antibodies can be prepared and, those monoclonal antibodies or the antigens, can be employed in well known antibody binding assays, diagnostic kits or tests to determine the presence or absence of particular HCMV antigen(s) and therefore the presence or absence of the virus or expression of the antigen(s) (in HCMV or other systems), or to determine whether an immune response to the virus or antigen(s) has simply been stimulated. Those monoclonal antibodies or the antigens can also be employed in immunoadsorption chromatography to recover or isolate HCMV or expression products of the inventive recombinant poxvirus.

More in particular, the inventive recombinants and compositions have numerous utilities, including:

(i) inducing an immunological response in seronegative individuals (use as or as part of a vaccine regimen);

(ii) therapy in seropositive individuals; and

(iii) a means for generating HCMV protein in vitro without the risk of virus infection.

The products of expression of the inventive recombinant poxvirus can be used directly to stimulate an immune response in either seronegative or seropositive individuals or in animals. Thus, the expression products can be used in compositions of the invention instead of or in addition to the inventive recombinant poxvirus.

Additionally, the inventive recombinant poxvirus and the expression products therefrom stimulate an immune or antibody response in humans and animals. From those antibodies, by techniques well-known in the art, monoclonal antibodies can be prepared and, those monoclonal antibodies or the expression products of the inventive poxvirus and composition can be employed in well known antibody binding assays, diagnostic kits or tests to determine the presence or absence of particular HCMV antigen(s) or antibody(ies) and therefore the presence or absence of the virus, or to determine whether an immune response to the virus or antigen(s) has simply been stimulated. Those monoclonal antibodies can also be employed in immunoadsorption chromatography to recover, isolate or detect HCMV or expression products of the inventive recombinant poxvirus. Methods for producing monoclonal antibodies and for uses of monoclonal antibodies, and, of uses and methods for HCMV antigens--the expression products of the inventive poxvirus and composition--are well known to those of ordinary skill in the art. They can be used in diagnostic methods, kits, tests or assays, as well as to recover materials by immunoadsorption chromatography or by immunoprecipitation.

Monoclonal antibodies are immunoglobulins produced by hybridoma cells. A monoclonal antibody reacts with a single antigenic determinant and provides greater specificity than a conventional, serum-derived antibody. Furthermore, screening a large number of monoclonal antibodies makes it possible to select an individual antibody with desired specificity, avidity and isotype. Hybridoma cell lines provide a constant, inexpensive source of chemically identical antibodies and preparations of such antibodies can be easily standardized. Methods for producing monoclonal antibodies are well known to those of ordinary skill in the art, e.g., Koprowski, H. et al., U.S. Pat. No. 4,196,265, issued Apr. 1, 1989, incorporated herein by reference.

Uses of monoclonal antibodies are known. One such use is in diagnostic methods, e.g., David, G. and Greene, H. U.S. Pat. No. 4,376,110, issued Mar. 8, 1983; incorporated herein by reference. Monoclonal antibodies have also been used to recover materials by immunoadsorption chromatography, e.g., Milstein, C. 1980, Scientific American 243:66, 70, incorporated herein by reference.

Furthermore, the inventive recombinant poxvirus or expression products therefrom can be used to stimulate a response in cells in vitro or ex vivo for subsequent reinfusion into a patient. If the patient is seronegative, the reinfusion is to stimulate an immune response, e.g., an immunological or antigenic response such as active immunization. In a seropositive individual, the reinfusion is to stimulate or boost the immune system against HCMV.

Accordingly, the inventive recombinant poxvirus has several utilities: In antigenic, immunological or vaccine compositions such as for administration to seronegative individuals. In therapeutic compositions in seropositive individuals in need of therapy to stimulate or boost the immune system against HCMV. In vitro to produce antigens which can be further used in antigenic, immunological or vaccine compositions or in therapeutic compositions. To generate antibodies (either by direct administration or by administration of an expression product of the inventive recombinant poxvirus) or expression products or antigens which can be further used: in diagnosis, tests or kits to ascertain the presence or absence of antigens in a sample such as sera, for instance, to ascertain the presence or absence of HCMV in a sample such as sera or, to determine whether an immune response has elicited to the virus or, to particular antigen(s); or, in immunoadsorption chromatography, immunoprecipitation and the like.

Furthermore, the recombinant poxviruses of the invention are useful for generating DNA for probes or for

PCR primers which can be used to detect the presence or absence of hybridizable DNA or to amplify DNA, e.g., to detect HCMV in a sample or for amplifying HCMV DNA.

Other utilities also exist for embodiments of the invention.

A better understanding of the present invention and of its many advantages will be had from the following examples, given by way of illustration.

EXAMPLES

DNA Cloning and Synthesis. Plasmids were constructed, screened and grown by standard procedures (Maniatis et al., 1982; Perkus et al., 1985; Piccini et al., 1987). Restriction endonucleases were obtained from Bethesda Research Laboratories, Gaithersburg, Md., New England Biolabs, Beverly, Mass.; and Boehringer Mannheim Biochemicals, Indianapolis, Ind. Klenow fragment of E. coli polymerase was obtained from Boehringer Mannheim Biochemicals. BAL-31 exonuclease and phage T4 DNA ligase were obtained from New England Biolabs. The reagents were used as specified by the various suppliers.

Synthetic oligodeoxyribonucleotides were prepared on a Biosearch 8750 or Applied Biosystems 380B DNA synthesizer as previously described (Perkus et al., 1989). DNA sequencing was performed by the dideoxy-chain termination method (Sanger et al., 1977) using Sequenase (Tabor et al., 1987) as previously described (Guo et al., 1989). DNA amplification by polymerase chain reaction (PCR) for sequence verification (Engelke et al., 1988) was performed using custom synthesized oligonucleotide primers and GeneAmp DNA amplification Reagent Kit (Perkin Elmer Cetus, Norwalk, Conn.) in an automated Perkin Elmer Cetus DNA Thermal Cycler. Excess DNA sequences were deleted from plasmids by restriction endonuclease digestion followed by limited digestion by BAL-31 exonuclease and mutagenesis (Mandecki, 1986) using synthetic oligonucleotides.

Cells, Virus, and Transfection. The origins and conditions of cultivation of the Copenhagen strain of vaccinia virus has been previously described (Guo et al., 1989). Generation of recombinant virus by recombination, in situ hybridization of nitrocellulose filters and screening for B-galactosidase activity are as previously described (Piccini et al., 1987).

The origins and conditions of cultivation of the Copenhagen strain of vaccinia virus and NYVAC has been previously described (Guo et al., 1989; Tartaglia et al., 1992). Generation of recombinant virus by recombination, in situ hybridization of nitrocellulose filters and screening for B-galactosidase activity are as previously described (Panicali et al., 1982; Perkus et al., 1989).

The parental canarypox virus (Rentschler strain) is a vaccinal strain for canaries. The vaccine strain was obtained from a wild type isolate and attenuated through more than 200 serial passages on chick embryo fibroblasts. A master viral seed was subjected to four successive plaque purifications under agar and one plaque clone was amplified through five additional passages after which the stock virus was used as the parental virus in in vitro recombination tests. The plaque purified canarypox isolate is designated ALVAC.

The strain of fowlpox virus (FPV) designated FP-1 has been described previously (Taylor et al., 1988a). It is an attenuated vaccine strain useful in vaccination of day old chickens. The parental virus strain Duvette was obtained in France as a fowlpox scab from a chicken. The virus was attenuated by approximately 50 serial passages in chicken embryonated eggs followed by 25 passages on chicken embryo fibroblast cells. The virus was subjected to four successive plaque purifications. One plaque isolate was further amplified in primary CEF cells and a stock virus, designated as TROVAC, established.

NYVAC, ALVAC and TROVAC viral vectors and their derivatives were propagated as described previously (Piccini et al., 1987; Taylor et al., 1988a,b). Vero cells and chick embryo fibroblasts (CEF) were propagated as described previously (Taylor et al., 1988a,b).

As to NYVAC and especially Examples 1 to 6, reference's made to U.S. Pat. No. 5,364,773, incorporated herein by reference.

Example 1 Construction of Plasmid pSD460 for Deletion of Thymidine Kinase Gene (J2R)

Referring now to FIG. 1, plasmid pSD406 contains vaccinia HindIII J (pos. 83359-88377) cloned into pUC8. pSD406 was cut with HindIII and PvuII, and the 1.7 kb fragment from the left side of HindIII J cloned into pUC8 cut with HindIII/SmaI, forming pSD447. pSD447 contains the entire gene for J2R (pos. 83855-84385). The initiation codon is contained within an NlaIII site and the termination codon is contained within an SspI site. Direction of transcription is indicated by an arrow in FIG. 1.

To obtain a left flanking arm, a 0.8 kb HindIII/EcoRI fragment was isolated from pSD447, then digested with NlaIII and a 0.5 kb HindIII/NlaIII fragment isolated. Annealed synthetic oligonucleotides MPSYN43/MPSYN44 (SEQ ID NO:1/SEQ ID NO:2)

                                 SmaI                                              MPSYN43 5'     TAATTAACTAGCTACCCGGG      3'                                    MPSYN44 3' GTACATTAATTGATCGATGGGCCCTTAA  5'                                              NlaIII                  EcoRI                                   

were ligated with the 0.5 kb HindIII/NlaIII fragment into pUC18 vector plasmid cut with HindIII/EcoRI, generating plasmid pSD449.

To obtain a restriction fragment containing a vaccinia right flanking arm and pUC vector sequences, pSD447 was cut with SspI (partial) within vaccinia sequences and HindIII at the pUC/vaccinia junction, and a 2.9 kb vector fragment isolated. This vector fragment was ligated with annealed synthetic oligonucleotides MPSYN45/MPSYN46 (SEQ ID NO:3/SEQ ID NO:4)

              HindIII SmaI               m         m                  NotI               SspI                                                                     MPSYN45 5'  AGCTTCCCGGGTAAGTAATACGTCAAGGAGAAAACGAAACGATCTGTAGTTAGCGGCCGCCT     AATTAACTAAT  3' MPSYN45                                                        MPSYN46 3'      AGGGCCCATTCATTATGCAGTTCCTCTTTTGCTT         TGCTAGACATCAATC     GCCGGCGGATTAATTGATTA  5' MPSYN46                                          

generating pSD459.

To combine the left and right flanking arms into one plasmid, a 0.5 kb HindIII/SmaI fragment was isolated from pSD449 and ligated with pSD459 vector plasmid cut with HindIII/SmaI, generating plasmid pSD460. pSD460 was used as donor plasmid for recombination with wild type parental vaccinia virus Copenhagen strain VC-2. ³² P labelled probe was synthesized by primer extension using MPSYN45 (SEQ ID NO:3) as template and the complementary 20 mer oligonucleotide MPSYN47 (SEQ ID NO:5) (5' TTAGTTAATTAGGCGGCCGC 3') as primer. Recombinant virus vP410 was identified by plaque hybridization.

Example 2 Construction of Plasmid pSD486 for Deletion of Hemorrhagic Region (B13r+B14r)

Referring now to FIG. 2, plasmid pSD419 contains vaccinia SalI G (pos. 160,744-173,351) cloned into pUC8. pSD422 contains the contiguous vaccinia SalI fragment to the right, SalI J (pos. 173,351-182,746) cloned into pUC8. To construct a plasmid deleted for the hemorrhagic region, u, B13R-B14R (pos. 172,549-173,552), pSD419 was used as the source for the left flanking arm and pSD422 was used as the source of the right flanking arm. The direction of transcription for the u region is indicated by an arrow in FIG. 2.

To remove unwanted sequences from pSD419, sequences to the left of the NcoI site (pos. 172,253) were removed by digestion of pSD419 with NcoI/SmaI followed by blunt ending with Klenow fragment of E. coli polymerase and ligation generating plasmid pSD476. A vaccinia right flanking arm was obtained by digestion of pSD422 with HpaI at the termination codon of B14R and by digestion with NruI 0.3 kb to the right. This 0.3 kb fragment was isolated and ligated with a 3.4 kb HincII vector fragment isolated from pSD476, generating plasmid pSD477. The location of the partial deletion of the vaccinia u region in pSD477 is indicated by a triangle. The remaining B13R coding sequences in pSD477 were removed by digestion with ClaI/HpaI, and the resulting vector fragment was ligated with annealed synthetic oligonucleotides SD22mer/SD20mer (SEQ ID NO:6/SEQ ID NO:7)

               ClaI         BamHI HpaI                                             SD22mer 5' CGATTACTATGAAGGATCCGTT  3'                                          SD20mer 3'   TAATGATACTTCCTAGGCAA  5'                                     

generating pSD479. pSD479 contains an initiation codon (underlined) followed by a BamHI site. To place E. coli Beta-galactosidase in the B13-B14 (u) deletion locus under the control of the u promoter, a 3.2 kb BamHI fragment containing the Beta-galactosidase gene (Shapira et al., 1983) was inserted into the BamHI site of pSD479, generating pSD479BG. pSD479BG was used as donor plasmid for recombination with vaccinia virus vP410. Recombinant vaccinia virus vP533 was isolated as a blue plaque in the presence of chromogenic substrate X-gal. In vP533 the B13R-B14R region is deleted and is replaced by Betagalactosidase.

To remove Beta-galactosidase sequences from vP533, plasmid pSD486, a derivative of pSD477 containing a polylinker region but no initiation codon at the u deletion junction, was utilized. First the ClaI/HpaI vector fragment from pSD477 referred to above was ligated with annealed synthetic oligonucleotides SD42mer/SD40mer (SEQ ID NO:8/SEQ ID NO:9)

                                            ClaI          SacI        XhoI                                          HpaI                                                                       SD42mer 5' CGATTACTAGATCTGAGCTCCCCGGGCTCGAGGG                                  ATCCGTT  3'                                                                    SD40mer 3'   TAATGATCTAGACTCGAGGGGCCCGAGCTCCC                                  TAGGCAA  5'                                                                                       BglII       SmaI                                            BamHI                                        

generating plasmid pSD478. Next the EcoRI site at the pUC/vaccinia junction was destroyed by digestion of pSD478 with EcoRI followed by blunt ending with Klenow fragment of E. coli polymerase and ligation, generating plasmid pSD478E⁻. pSD478E⁻ was digested with BamHI and HpAI and ligated with annealed synthetic oligonucleotides HEM5/HEM6 (SEQ ID NO:10/SEQ ID NO:11)

                   BamHI EcoRI   HpaI                                                      HEM5 5'  GATCCGAATTCTAGCT  3'                                                  HEM6 3'      GCTTAAGATCGA  5'                                     

generating plasmid pSD486. pSD486 was used as donor plasmid for recombination with recombinant vaccinia virus vP533, generating vP553, which was isolated as a clear plaque in the presence of X-gal.

Example 3 Construction of Plasmid pMP494Δ for Deletion of ATI Region (A26L)

Referring now to FIG. 3, pSD414 contains SalI B cloned into pUC8. To remove unwanted DNA sequences to the left of the A26L region, pSD414 was cut with XbaI within vaccinia sequences (pos. 137,079) and with HindIII at the pUC/vaccinia junction, then blunt ended with Klenow fragment of E. coli polymerase and ligated, resulting in plasmid pSD483. To remove unwanted vaccinia DNA sequences to the right of the A26L region, pSD483 was cut with EcoRI (pos. 140,665 and at the pUC/vaccinia junction) and ligated, forming plasmid pSD484. To remove the A26L coding region, pSD484 was cut with NdeI (partial) slightly upstream from the A26L ORF (pos. 139,004) and with HpaI (pos. 137,889) slightly downstream from the A26L ORF. The 5.2 kb vector fragment was isolated and ligated with annealed synthetic oligonucleotides ATI3/ATI4 (SEQ ID NO:12/SEQ ID NO:13)

           NdeI                                                         BglII      EcoRI HpaI                                                                     ATI3 5' TATGAGTAACTTAACTCTTTTGTTAATTAAAAGTATATTCAAAAAATAAGTTATATAAATAGATCT     GAATTCGTT  3' ATI3                                                             ATI4 3'   ACTCATTGAATTGAGAAAACAATTAATTTTCATATAAGTTTTTTATTCAATATATTTATCTAGA     CTTAAGCAA  5' ATI4                                                        

reconstructing the region upstream from A26L and replacing the A26L ORF with a short polylinker region containing the restriction sites BQlII, EcoRI and HpaI, as indicated above. The resulting plasmid was designated pSD485. Since the BqlII and EcoRI sites in the polylinker region of pSD485 are not unique, unwanted BqlII and EcoRI sites were removed from plasmid pSD483 (described above) by digestion with BglII (pos. 140,136) and with EcoRI at the pUc/vaccinia junction, followed by blunt ending with Klenow fragment of E. coli polymerase and ligation. The resulting plasmid was designated pSD489. The 1.8 kb ClaI (pos. 137,198)/EcoRV (pos. 139,048) fragment from pSD489 containing the A26L ORF was replaced with the corresponding 0.7 kb polylinker-containing CaI/EcoRV fragment from pSD485, generating pSD492. The BqlII and EcoRI sites in the polylinker region of pSD492 are unique.

A 3.3 kb BglII cassette containing the E. coli Beta-galactosidase gene (Shapira et al., 1983) under the control of the vaccinia 11 kDa promoter (Bertholet et al., 1985; Perkus et al., 1990) was inserted into the BglII site of pSD492, forming pSD493KBG. Plasmid pSD493KBG was used in recombination with rescuing virus vP553. Recombinant vaccinia virus, vP581, containing Beta-galactosidase in the A26L deletion region, was isolated as a blue plaque in the presence of X-gal.

To generate a plasmid for the removal of Beta-galactosidase sequences from vaccinia recombinant virus vP581, the polylinker region of plasmid pSD492 was deleted by mutagenesis (Mandecki, 1986) using synthetic oligonucleotide MPSYN177 (SEQ ID NO:14)

(5' AAAATGGGCGTGGATTGTTAACTTTATATAACTTATTTTTTGAATATAC 3'). In the resulting plasmid, pMP494Δ, vaccinia DNA encompassing positions [137,889-138,937], including the entire A26L ORF is deleted. Recombination between the pMP494Δ and the Beta-galactosidase containing vaccinia recombinant, vP581, resulted in vaccinia deletion mutant vP618, which was isolated as a clear plaque in the presence of X-gal.

Example 4 Construction of Plasmid pSD467 for Deletion of Hemagglutinin Gene (A56R)

Referring now to FIG. 4, vaccinia SalI G restriction fragment (pos. 160,744-173,351) crosses the HindIII A/B junction (pos. 162,539). pSD419 contains vaccinia SalI G cloned into pUC8. The direction of transcription for the hemagglutinin (HA) gene is indicated by an arrow in FIG. 4. Vaccinia sequences derived from HindIII B were removed by digestion of pSD419 with HindIII within vaccinia sequences and at the pUC/vaccinia junction followed by ligation. The resulting plasmid, pSD456, contains the HA gene, A56R, flanked by 0.4 kb of vaccinia sequences to the left and 0.4 kb of vaccinia sequences to the right. A56R coding sequences were removed by cutting pSD456 with RsaI (partial; pos. 161,090) upstream from A56R coding sequences, and with EagI (pos. 162,054) near the end of the gene. The 3.6 kb RsaI/EagI vector fragment from pSD456 was isolated and ligated with annealed synthetic oligonucleotides MPSYN59 (SEQ ID NO:15), MPSYN62 (SEQ ID NO:16), MPSYN60 (SEQ ID NO:17), and MPSYN61 (SEQ ID NO:18)

            RsaI                                                                   MPSYN595'ACACGAATGATTTTCTAAAGTATTTGGAAAGTTTTATAGGT-                            MPSYN623'TGTGCTTACTAAAAGATTTCATAAACCTTTCAAAATATCCA-                            MPSYN59   AGTTGATAGAACAAAATACATAATTT 3'                                        MPSYN62   TCAACTATCT 5'                                                        MPSYN605'               TGTAAAAATAAATCACTTTTTATA-                              MPSYN613'TGTTTTATGTATTAAAACATTTTTATTTAGTGAAAAATAT-                                          BglII SmaI  PstI  EagI                                            MPSYN60   CTAAGATCTCCCGGGCTGCAGC      3'                                       MPSYN61   GATTCTAGAGGGCCCGACGTCGCCGG  5'                                  

reconstructing the DNA sequences upstream from the A56R ORF and replacing the A56R ORF with a polylinker region as indicated above. The resulting plasmid is pSD466. The vaccinia deletion in pSD466 encompasses positions [161,185-162,053]. The site of the deletion in pSD466 is indicated by a triangle in FIG. 4.

A 3.2 kb BqlII/BamHI (partial) cassette containing the E. coli Beta-galactosidase gene (Shapira et al., 1983) under the control of the vaccinia 11 kDa promoter (Bertholet et al., 1985; Guo et al., 1989) was inserted into the BglII site of pSD466, forming pSD466KBG. Plasmid pSD466KBG was used in recombination with rescuing virus vP618. Recombinant vaccinia virus, vP708, containing Beta-galactosidase in the A56R deletion, was isolated as a blue plaque in the presence of X-gal.

Beta-galactosidase sequences were deleted from vP708 using donor plasmid pSD467. pSD467 is identical to pSD466, except that EcoRI, SmaI and BamHI sites were removed from the pUC/vaccinia junction by digestion of pSD466 with EcoRI/BamHI followed by blunt ending with Klenow fragment of E. coli polymerase and ligation. Recombination between vP708 and pSD467 resulted in recombinant vaccinia deletion mutant, vP723, which was isolated as a clear plaque in the presence of X-gal.

Example 5 Construction of Plasmid pMPCSK1Δ for Deletion of Open Reading Frames [C7L-K1L]

Referring now to FIG. 5, the following vaccinia clones were utilized in the construction of pMPCSK1Δ. pSD420 is SalI H cloned into pUC8. pSD435 is KpnI F cloned into pUC18. pSD435 was cut with SphI and religated, forming pSD451. In pSD451, DNA sequences to the left of the SphI site (pos. 27,416) in HindIII M are removed (Perkus et al., 1990). pSD409 is HindIII M cloned into pUC8.

To provide a substrate for the deletion of the [C7L-K1L] gene cluster from vaccinia, E. coli Beta-galactosidase was first inserted into the vaccinia M2L deletion locus (Guo et al., 1990) as follows. To eliminate the BglII site in pSD409, the plasmid was cut with BglII in vaccinia sequences (pos. 28,212) and with BamHI at the pUC/vaccinia junction, then ligated to form plasmid pMP409B. pMP409B was cut at the unique SphI site (pos. 27,416). M2L coding sequences were removed by mutagenesis (Guo et al., 1990; Mandecki, 1986) using synthetic oligonucleotide MPSYN82 (SEQ. ID NO:19)

                              BglII                                                5' TTTCTGTATATTTGCACCAATTTAGATCTTACTCAAAATATGTAACAATA 3'                  

The resulting plasmid, pMP409D, contains a unique BglII site inserted into the M2L deletion locus as indicated above. A 3.2 kb BamHI (partial)/BglII cassette containing the E. coli Beta-galactosidase gene (Shapira et al., 1983) under the control of the 11 kDa promoter (Bertholet et al., 1985) was inserted into pMP409D cut with BglII. The resulting plasmid, pMP409DBG (Guo et al., 1990), was used as donor plasmid for recombination with rescuing vaccinia virus vP723. Recombinant vaccinia virus, vP784, containing Beta-galactosidase inserted into the M2L deletion locus, was isolated as a blue plaque in the presence of X-gal.

A plasmid deleted for vaccinia genes [C7L-K1L] was assembled in pUC8 cut with SmaI, HindIII and blunt ended with Klenow fragment of E. coli polymerase. The left flanking arm consisting of vaccinia HindIII C sequences was obtained by digestion of pSD420 with XbaI (pos. 18,628) followed by blunt ending with Klenow fragment of E. coli polymerase and digestion with BglII (pos. 19,706). The right flanking arm consisting of vaccinia HindIII K sequences was obtained by digestion of pSD451 with BglII (pos. 29,062) and EcoRV (pos. 29,778). The resulting plasmid, pMP581CK is deleted for vaccinia sequences between the BglII site (pos. 19,706) in HindIII C and the BglII site (pos. 29,062) in HindIII K. The site of the deletion of vaccinia sequences in plasmid pMP581CK is indicated by a triangle in FIG. 5.

To remove excess DNA at the vaccinia deletion junction, plasmid pMP581CK, was cut at the NcoI sites within vaccinia sequences (pos. 18,811; 19,655), treated with Bal-31 exonuclease and subjected to mutagenesis (Mandecki, 1986) using synthetic oligonucleotide MPSYN233 (SEQ ID NO:20) 5'-TGTCATTTAACACTATACTCATATTAATAAAAATAATATTTATT-3'. The resulting plasmid, pMPCSK1Δ, is deleted for vaccinia sequences positions 18,805-29,108, encompassing 12 vaccinia open reading frames [C7L-K1L]. Recombination between pMPCSK1Δ and the Beta-galactosidase containing vaccinia recombinant, vP784, resulted in vaccinia deletion mutant, vP804, which was isolated as a clear plaque in the presence of X-gal.

Example 6 Construction of Plasmid pSD548 for Deletion of Large Subunit, Ribonucleotide Reductase (I4L)

Referring now to FIG. 6, plasmid pSD405 contains vaccinia HindIII I (pos. 63,875-70,367) cloned in pUC8. pSD405 was digested with EcoRV within vaccinia sequences (pos. 67,933) and with SmaI at the pUC/vaccinia junction, and ligated, forming plasmid pSD518. pSD518 was used as the source of all the vaccinia restriction fragments used in the construction of pSD548.

The vaccinia I4L gene extends from position 67,371-65,059. Direction of transcription for I4L is indicated by an arrow in FIG. 6. To obtain a vector plasmid fragment deleted for a portion of the I4L coding sequences, pSD518 was digested with BamHI (pos. 65,381) and HpaI (pos. 67,001) and blunt ended using Klenow fragment of E. coli polymerase. This 4.8 kb vector fragment was ligated with a 3.2 kb SmaI cassette containing the E. coli Beta-galactosidase gene (Shapira et al., 1983) under the control of the vaccinia 11 kDa promoter (Bertholet et al., 1985; Perkus et al., 1990), resulting in plasmid pSD524KBG. pSD524KBG was used as donor plasmid for recombination with vaccinia virus vP804. Recombinant vaccinia virus, vP855, containing Beta-galactosidase in a partial deletion of the I4L gene, was isolated as a blue plaque in the presence of X-gal.

To delete Beta-galactosidase and the remainder of the I4L ORF from vP855, deletion plasmid pSD548 was constructed. The left and right vaccinia flanking arms were assembled separately in pUC8 as detailed below and presented schematically in FIG. 6.

To construct a vector plasmid to accept the left vaccinia flanking arm, pUC8 was cut with BamHI/EcoRI and ligated with annealed synthetic oligonucleotides 518A1/518A2 (SEQ ID NO:21/SEQ ID NO:22)

             BamHI   RsaI                                                          BglI    EcoRI                                                                  518A1 5' GATCCTGAGTACTTTGTAATATAATGATATATATTTTCACTTTATCTCATTTGAGAATAAAAAGA     TCTTAGG     3'   518A1                                                         518A2 3'     GACTCATGAAACATTATATTACTATATATAAAAGTGAAATAGAGTAAACTCTTATTTTTCT     AGAATCCTTAA 5'   518A2                                                    

forming plasmid pSD531. pSD531 was cut with RsaI (partial) and BamHI and a 2.7 kb vector fragment isolated. pSD518 was cut with BglII (pos. 64,459)/RsaI (pos. 64,994) and a 0.5 kb fragment isolated. The two fragments were ligated together, forming pSD537, which contains the complete vaccinia flanking arm left of the I4L coding sequences.

To construct a vector plasmid to accept the right vaccinia flanking arm, pUC8 was cut with BamHI/EcoRI and ligated with annealed synthetic oligonucleotides 518B1/518B2 (SEQ ID NO:23/SEQ ID NO:24)

               BamHI BglII SmaI                                                      RsaI   EcoRI                                                                 518B1   5'  GATCCAGATCTCCCGGGAAAAAAATTATTTAACTTTTCATTAATAGGGATTTGACGTATGTA     GCGTACTAGG     3'   518B1                                                      518B2   3'      GTCTAGAGGGCCCTTTTTTTAATAAATTGAAAAGTAATTATCCCTAAACTGCATACTA     CGCATGATCCTTAA 5'   518B2                                                 

forming plasmid pSD532. pSD532 was cut with RsaI (partial)/EcoRI and a 2.7 kb vector fragment isolated. pSD518 was cut with RsaI within vaccinia sequences (pos. 67,436) and EcoRI at the vaccinia/pUC junction, and a 0.6 kb fragment isolated. The two fragments were ligated together, forming pSD538, which contains the complete vaccinia flanking arm to the right of I4L coding sequences.

The right vaccinia flanking arm was isolated as a 0.6 kb EcoRI/BglII fragment from pSD538 and ligated into pSD537 vector plasmid cut with EcoRI/BglII. In the resulting plasmid, pSD539, the I4L ORF (pos. 65,047-67,386) is replaced by a polylinker region, which is flanked by 0.6 kb vaccinia DNA to the left and 0.6 kb vaccinia DNA to the right, all in a pUC background. The site of deletion within vaccinia sequences is indicated by a triangle in FIG. 6. To avoid possible recombination of Beta-galactosidase sequences in the pUC-derived portion of pSD539 with Beta-galactosidase sequences in recombinant vaccinia virus vP855, the vaccinia I4L deletion cassette was moved from pSD539 into pRC11, a pUC derivative from which all Beta-galactosidase sequences have been removed and replaced with a polylinker region (Colinas et al., 1990). pSD539 was cut with EcoRI/PstI and the 1.2 kb fragment isolated. This fragment was ligated into pRC11 cut with EcoRI/PstI (2.35 kb), forming pSD548. Recombination between pSD548 and the Beta-galactosidase containing vaccinia recombinant, vP855, resulted in vaccinia deletion mutant vP866, which was isolated as a clear plaque in the presence of X-gal.

DNA from recombinant vaccinia virus vP866 was analyzed by restriction digests followed by electrophoresis on an agarose gel. The restriction patterns were as expected. Polymerase chain reactions (PCR) (Engelke et al., 1988) using vP866 as template and primers flanking the six deletion loci detailed above produced DNA fragments of the expected sizes. Sequence analysis of the PCR generated fragments around the areas of the deletion junctions confirmed that the junctions were as expected. Recombinant vaccinia virus vP866, containing the six engineered deletions as described above, was designated vaccinia vaccine strain "NYVAC."

Example 7 Insertion of a Rabies Glycoprotein G Gene Into NYVAC

The gene encoding rabies glycoprotein G under the control of the vaccinia H6 promoter (Taylor et al., 1988a,b) was inserted into TK deletion plasmid pSD513. pSD513 is identical to plasmid pSD460 (FIG. 1) except for the presence of a polylinker region.

Referring now to FIG. 7, the polylinker region was inserted by cutting pSD460 with SmaI and ligating the plasmid vector with annealed synthetic oligonucleotides VQ1A/VQ1B (SEQ ID NO:25/SEQ ID NO:26)

           SmaI BglII XhoI  PstI  NarI  BamHI                                      VQ1A 5' GGGAGATCTCTCGAGCTGCAGGGCGCCGGATCCTTTTTCT 3'                            VQ1B 3' CCCTCTAGAGAGCTCGACGTCCCGCGGCCTAGGAAAAAGA 5'                       

to form vector plasmid pSD513. pSD513 was cut with SmaI and ligated with a SmaI ended 1.8 kb cassette containing the gene encoding the rabies glycoprotein G gene under the control of the vaccinia H6 promoter (Taylor et al., 1988a,b). The resulting plasmid was designated pRW842. pRW842 was used as donor plasmid for recombination with NYVAC rescuing virus (vP866). Recombinant vaccinia virus vP879 was identified by plaque hybridization using ³² P labelled DNA probe to rabies glycoprotein G coding sequences.

The modified recombinant viruses of the present invention provide advantages as recombinant vaccine vectors. The attenuated virulence of the vector advantageously reduces the opportunity for the possibility of a runaway infection due to vaccination in the vaccinated individual and also diminishes transmission from vaccinated to unvaccinated individuals or contamination of the environment.

The modified recombinant viruses are also advantageously used in a method for expressing a gene product in a cell cultured in vitro by introducing into the cell the modified recombinant virus having foreign DNA which codes for and expresses gene products in the cell.

Example 8 Construction of ALVAC Recombinants Expressing Rabies Virus Glycoprotein G

This example describes the development of ALVAC, a canarypox virus vector and, of a canarypox-rabies recombinant designated as ALVAC-RG (vCP65) and its safety and efficacy.

Cells and Viruses. The parental canarypox virus (Rentschler strain) is a vaccinal strain for canaries. The vaccine strain was obtained from a wild type isolate and attenuated through more than 200 serial passages on chick embryo fibroblasts. A master viral seed was subjected to four successive plaque purifications under agar and one plaque clone was amplified through five additional passages after which the stock virus was used as the parental virus in in vitro recombination tests. The plaque purified canarypox isolate is designated ALVAC.

Construction of a Canarypox Insertion Vector. An 880 bp canarypox PvuII fragment was cloned between the PvuII sites of pUC9 to form pRW764.5. The sequence of this fragment is shown in FIG. 8 (SEQ ID NO:27) between positions 1372 and 2251. The limits of an open reading frame designated as C5 were defined. It was determined that the open reading frame was initiated at position 166 within the fragment and terminated at position 487. The C5 deletion was made without interruption of open reading frames. Bases from position 167 through position 455 were replaced with the sequence (SEQ ID NO:28) GCTTCCCGGGAATTCTAGCTAGCTAGTTT. This replacement sequence contains HindIII, SmaI and EcoRI insertion sites followed by translation stops and a transcription termination signal recognized by vaccinia virus RNA polymerase (Yuen et al., 1987). Deletion of the C5 ORF was performed as described below. Plasmid pRW764.5 was partially cut with RsaI and the linear product was isolated. The RsaI linear fragment was recut with BglII and the pRW764.5 fragment now with a RsaI to BglII deletion from position 156 to position 462 was isolated and used as a vector for the following synthetic oligonucleotides:

RW145 (SEQ ID NO:29):

ACTCTCAAAAGCTTCCCGGGAATTCTAGCTAGCTAGTTTTTATAAA

RW146 (SEQ ID NO:30):

GATCTTTATAAAAACTAGCTAGCTAGAATTCCCGGGAAGCTTTTGAGAGT

Oligonucleotides RW145 and RW146 were annealed and inserted into the pRW 764.5 RsaI and BglII vector described above. The resulting plasmid is designated pRW831.

Construction of Insertion Vector Containing the Rabies G Gene. Construction of pRW838 is illustrated below. oligonucleotides A through E, which overlap the translation initiation codon of the H6 promoter with the ATG of rabies G, were cloned into pUC9 as pRW737. Oligonucleotides A through E contain the H6 promoter, starting at NruI, through the HindIII site of rabies G followed by BglII. Sequences of oligonucleotides A through E ((SEQ ID NO:31)-(SEQ ID NO:35)) are:

    A (SEQ ID NO:31):CTGAAATTATTTCATTATCGCGATATCCGTTAA                                       GTTTGTATCGTAATGGTTCCTCAGGCTCTCCTGTTTGT                               B (SEQ ID NO:32):CATTACGATACAAACTTAACGGATATCGCGATAA                                      TGAAATAATTTCAG                                                       C (SEQ ID NO:33: ACCCCTTCTGGTTTTTCCGTTGTGTTTTGGGAAA                                      TTCCCTATTTACACGATCCCAGACAAGCTTAGATCTCAG                              D (SEQ ID NO:34):CTGAGATCTAAGCTTGTCTGGGATCGTGTAAATA                                      GGGAATTTCCCAAAACA                                                    E (SEQ ID NO:35):CAACGGAAAAACCAGAAGGGGTACAAACAGGAGA                                      GCCTGAGGAAC                                                     

The diagram of annealed oligonucleotides A through E is as follows: ##STR1##

Oligonucleotides A through E were kinased, annealed (95° C. for 5 minutes, then cooled to room temperature), and inserted between the PvuII sites of pUC9. The resulting plasmid, pRW737, was cut with HindIII and BglII and used as a vector for the 1.6 kbp HindIII-BglII fragment of ptg155PRO (Kieny et al., 1984) generating pRW739. The ptg155PRO HindIII site is 86 bp downstream of the rabies G translation initiation codon. BglII is downstream of the rabies G translation stop codon in ptg155PRO. pRW739 was partially cut with NruI, completely cut with BglII, and a 1.7 kbp NruI-BglII fragment, containing the 3' end of the H6 promoter previously described (Taylor et al., 1988a,b; Guo et al., 1989; Perkus et al., 1989) through the entire rabies G gene, was inserted between the NruI and BamHI sites of pRW824. The resulting plasmid is designated pRW832. Insertion into pRW824 added the H6 promoter 5' of NruI. The pRW824 sequence of BamHI followed by SmaI is (SEQ ID NO:36): GGATCCCCGGG. pRW824 is a plasmid that contains a nonpertinent gene linked precisely to the vaccinia virus H6 promoter. Digestion with NruI and BamHI completely excised this nonpertinent gene. The 1.8 kbp pRW832 SmaI fragment, containing H6 promoted rabies G, was inserted into the SmaI of pRW831, to form plasmid pRW838.

Development of ALVAC-RG. Plasmid pRW838 was transfected into ALVAC infected primary CEF cells by using the calcium phosphate precipitation method previously described (Panicali et al., 1982; Piccini et al., 1987). Positive plaques were selected on the basis of hybridization to a specific rabies G probe and subjected to 6 sequential rounds of plaque purification until a pure population was achieved. One representative plaque was then amplified and the resulting ALVAC recombinant was designated ALVAC-RG (vCP65) (see also FIGS. 9A and 9B). The correct insertion of the rabies G gene into the ALVAC genome without subsequent mutation was confirmed by sequence analysis.

Immunofluorescence. During the final stages of assembly of mature rabies virus particles, the glycoprotein component is transported from the golgi apparatus to the plasma membrane where it accumulates with the carboxy terminus extending into the cytoplasm and the bulk of the protein on the external surface of the cell membrane. In order to confirm that the rabies glycoprotein expressed in ALVAC-RG was correctly presented, immunofluorescence was performed on primary CEF cells infected with ALVAC or ALVAC-RG. Immunofluorescence was performed as previously described (Taylor et al., 1990) using a rabies G monoclonal antibody. Strong surface fluorescence was detected on CEF cells infected with ALVAC-RG but not with the parental ALVAC.

Immunoprecipitation. Preformed monolayers of primary CEF, Vero (a line of African Green monkey kidney cells ATCC # CCL81) and MRC-5 cells (a fibroblast-like cell line derived from normal human fetal lung tissue ATCC # CCL171) were inoculated at 10 pfu per cell with parental virus ALVAC and recombinant virus ALVAC-RG in the presence of radiolabelled ³⁵ S-methionine and treated as previously described (Taylor et al., 1990). Immunoprecipitation reactions were performed using a rabies G specific monoclonal antibody. Efficient expression of a rabies specific glycoprotein with a molecular weight of approximately 67 kDa was detected with the recombinant ALVAC-RG. No rabies specific products were detected in uninfected cells or cells infected with the parental ALVAC virus.

Sequential Passaging Experiment. In studies with ALVAC virus in a range of non-avian species no proliferative infection or overt disease was observed (Taylor et al., 1991b). However, in order to establish that neither the parental nor recombinant virus could be adapted to grow in non-avian cells, a sequential passaging experiment was performed.

The two viruses, ALVAC and ALVAC-RG, were inoculated in 10 sequential blind passages in three cell substrates:

(1) Primary chick embryo fibroblast (CEF) cells produced from 11 day old white leghorn embryos;

(2) Vero cells--a continuous line of African Green monkey kidney cells (ATCC # CCL81); and

(3) MRC-5 cells--a diploid cell line derived from human fetal lung tissue (ATCC # CCL171).

The initial inoculation was performed at an m.o.i. of 0.1 pfu per cell using three 60 mm dishes of each cell substrate containing 2×10⁶ cells per dish. One dish was inoculated in the presence of 40 μg/ml of Cytosine arabinoside (Ara C), an inhibitor of DNA replication. After an absorption period of 1 hour at 37° C., the inoculum was removed and the monolayer washed to remove unabsorbed virus. At this time the medium was replaced with 5 ml of EMEM+2% NBCS on two dishes (samples t0 and t7) and 5ml of EMEM+2% NBCS containing 40 μg/ml Ara C on the third (sample t7A). Sample to was frozen at -70° C. to provide an indication of the residual input virus. Samples t7 and t7A were incubated at 37° C. for 7 days, after which time the contents were harvested and the cells disrupted by indirect sonication.

One ml of sample t7 of each cell substrate was inoculated undiluted onto three dishes of the same cell substrate (to provide samples t0, t7 and t7A) and onto one dish of primary CEF cells. Samples to, t7 and t7A were treated as for passage one. The additional inoculation on CEF cells was included to provide an amplification step for more sensitive detection of virus which might be present in the non-avian cells.

This procedure was repeated for 10 (CEF and MRC-5) or 8 (Vero) sequential blind passages. Samples were then frozen and thawed three times and assayed by titration on primary CEF monolayers.

Virus yield in each sample was then determined by plaque titration on CEF monolayers under agarose. Summarized results of the experiment are shown in Tables 1 and 2.

The results indicate that both the parental ALVAC and the recombinant ALVAC-RG are capable of sustained replication on CEF monolayers with no loss of titer. In Vero cells, levels of virus fell below the level of detection after 2 passages for ALVAC and 1 passage for ALVAC-RG. In MRC-5 cells, a similar result was evident, and no virus was detected after 1 passage. Although the results for only four passages are shown in Tables 1 and 2 the series was continued for 8 (Vero) and 10 (MRC-5) passages with no detectable adaptation of either virus to growth in the non-avian cells.

In passage 1 relatively high levels of virus were present in the t7 sample in MRC-5 and Vero cells. However this level of virus was equivalent to that seen in the to sample and the t7A sample incubated in the presence of Cytosine arabinoside in which no viral replication can occur. This demonstrated that the levels of virus seen at 7 days in non-avian cells represented residual virus and not newly replicated virus.

In order to make the assay more sensitive, a portion of the 7 day harvest from each cell substrate was inoculated onto a permissive CEF monolayer and harvested at cytopathic effect (CPE) or at 7 days if no CPE was evident. The results of this experiment are shown in Table 3. Even after amplification through a permissive cell substrate, virus was only detected in MRC-5 and Vero cells for two additional passages. These results indicated that under the conditions used, there was no adaptation of either virus to growth in Vero or MRC-5 cells.

Inoculation of Macaques. Four HIV seropositive macaques were initially inoculated with ALVAC-RG as described in Table 4. After 100 days these animals were re-inoculated to determine a booster effect, and an additional seven animals were inoculated with a range of doses. Blood was drawn at appropriate intervals and sera analyzed, after heat inactivation at 56° C. for 30 minutes, for the presence of anti-rabies antibody using the Rapid Fluorescent Focus Inhibition Assay (Smith et al., 1973).

Inoculation of Chimpanzees. Two adult male chimpanzees (50 to 65 kg weight range) were inoculated intramuscularly or subcutaneously with 1×10⁷ pfu of vCP65. Animals were monitored for reactions and bled at regular intervals for analysis for the presence of anti-rabies antibody with the RFFI test (Smith et al., 1973). Animals were re-inoculated with an equivalent dose 13 weeks after the initial inoculation.

Inoculation of Mice. Groups of mice were inoculated with 50 to 100 μl of a range of dilutions of different batches of vCP65. Mice were inoculated in the footpad. On day 14, mice were challenged by intracranial inoculation of from 15 to 43 mouse LD₅₀ of the virulent CVS strain of rabies virus. Survival of mice was monitored and a protective dose 50% (PD₅₀) calculated at 28 days post-inoculation.

Inoculation of Dogs and Cats. Ten beagle dogs, 5 months old, and 10 cats, 4 months old, were inoculated subcutaneously with either 6.7 or 7.7 log₁₀ TCID₅₀ of ALVAC-RG. Four dogs and four cats were not inoculated. Animals were bled at 14 and 28 days post-inoculation and anti-rabies antibody assessed in an RFFI test. The animals receiving 6.7 log₁₀ TCID₅₀ of ALVAC-RG were challenged at 29 days post-vaccination with 3.7 log₁₀ mouse LDSO (dogs) or 4.3 log₁₀ mouse LD₅₀ (cats) of the NYGS rabies virus challenge strain.

Inoculation of Squirrel Monkeys. Three groups of four squirrel monkeys (Saimiri sciureus) were inoculated with one of three viruses (a) ALVAC, the parental canarypox virus, (b) ALVAC-RG, the recombinant expressing the rabies G glycoprotein or (c) vCP37, a canarypox recombinant expressing the envelope glycoprotein of feline leukemia virus. Inoculations were performed under ketamine anaesthesia. Each animal received at the same time: (1) 20 μl instilled on the surface of the right eye without scarification; (2) 100 μl as several droplets in the mouth; (3) 100 μl in each of two intradermal injection sites in the shaven skin of the external face of the right arm; and (4) 100 μl in the anterior muscle of the right thigh.

Four monkeys were inoculated with each virus, two with a total of 5.0 log₁₀ pfu and two with a total of 7.0 log₁₀ pfu. Animals were bled at regular intervals and sera analyzed for the presence of antirabies antibody using an RFFI test (Smith et al., 1973). Animals were monitored daily for reactions to vaccination. Six months after the initial inoculation the four monkeys receiving ALVAC-RG, two monkeys initially receiving vCP37, and two monkeys initially receiving ALVAC, as well as one naive monkey were inoculated with 6.5 log₁₀ pfu of ALVAC-RG subcutaneously. Sera were monitored for the presence of rabies neutralizing antibody in an RFFI test (Smith et al., 1973).

Inoculation of Human Cell Lines with ALVAC-RG. In order to determine whether efficient expression of a foreign gene could be obtained in non-avian cells in which the virus does not productively replicate, five cell types, one avian and four non-avian, were analyzed for virus yield, expression of the foreign rabies G gene and viral specific DNA accumulation. The cells inoculated were:

(a) Vero, African Green monkey kidney cells, ATCC# CCL81;

(b) MRC-5, human embryonic lung, ATCC # CCL 171;

(c) WISH human amnion, ATCC # CCL 25;

(d) Detroit-532, human foreskin, Downs's syndrome, ATCC # CCL 54; and

(e) Primary CEF cells.

Chicken embryo fibroblast cells produced from 11 day old white leghorn embryos were included as a positive control. All inoculations were performed on preformed monolayers of 2×10⁶ cells as discussed below.

A. Methods for DNA analysis.

Three dishes of each cell line were inoculated at 5 pfu/cell of the virus under test, allowing one extra dish of each cell line un-inoculated. One dish was incubated in the presence of 40 μg/ml of cytosine arabinoside (Ara C). After an adsorption period of 60 minutes at 37° C., the inoculum was removed and the monolayer washed twice to remove unadsorbed virus. Medium (with or without Ara C) was then replaced. Cells from one dish (without Ara C) were harvested as a time zero sample. The remaining dishes were incubated at 37° C. for 72 hours, at which time the cells were harvested and used to analyze DNA accumulation. Each sample of 2×10⁶ cells was resuspended in 0.5 ml phosphate buffered saline (PBS) containing 40 mM EDTA and incubated for 5 minutes at 37° C. An equal volume of 1.5% agarose prewarmed at 42° C. and containing 120 mM EDTA was added to the cell suspension and gently mixed. The suspension was transferred to an agarose plug mold and allowed to harden for at least 15 min. The agarose plugs were then removed and incubated for 12-16 hours at 50° C. in a volume of lysis buffer (1% sarkosyl, 100 μg/ml proteinase K, 10 mM Tris HCl pH 7.5, 200 mM EDTA) that completely covers the plug. The lysis buffer was then replaced with 5.0 ml sterile 0.5×TBE (44.5 mM Tris-borate, 44.5 mM boric acid, 0.5 mM EDTA) and equilibrated at 4° C. for 6 hours with 3 changes of TBE buffer. The viral DNA within the plug was fractionated from cellular RNA and DNA using a pulse field electrophoresis system. Electrophoresis was performed for 20 hours at 180 V with a ramp of 50-90 sec at 15° C. in 0.5×TBE. The DNA was run with lambda DNA molecular weight standards. After electrophoresis the viral DNA band was visualized by staining with ethidium bromide. The DNA was then transferred to a nitrocellulose membrane and probed with a radiolabelled probe prepared from purified ALVAC genomic DNA.

B. Estimation of virus yield.

Dishes were inoculated exactly as described above, with the exception that input multiplicity was 0.1 pfu/cell. At 72 hours post infection, cells were lysed by three successive cycles of freezing and thawing. Virus yield was assessed by plaque titration on CEF monolayers.

C. Analysis of expression of Rabies G gene.

Dishes were inoculated with recombinant or parental virus at a multiplicity of 10 pfu/cell, allowing an additional dish as an uninfected virus control. After a one hour absorption period, the medium was removed and replaced with methionine free medium. After a 30 minute period, this medium was replaced with methionine-free medium containing 25 uCi/ml of ³⁵ S-Methionine. Infected cells were labelled overnight (approximately 16 hours), then lysed by the addition of buffer A lysis buffer. Immunoprecipitation was performed as previously described (Taylor et al., 1990) using a rabies G specific monoclonal antibody.

Results: Estimation of Viral Yield. The results of titration for yield at 72 hours after inoculation at 0.1 pfu per cell are shown in Table 5. The results indicate that while a productive infection can be attained in the avian cells, no increase in virus yield can be detected by this method in the four non-avian cell systems.

Analysis of Viral DNA Accumulation. In order to determine whether the block to productive viral replication in the non-avian cells occurred before or after DNA replication, DNA from the cell lysates was fractionated by electrophoresis, transferred to nitrocellulose and probed for the presence of viral specific DNA. DNA from uninfected CEF cells, ALVAC-RG infected CEF cells at time zero, ALVACRG infected CEF cells at 72 hours post-infection and ALVACRG infected CEF cells at 72 hours post-infection in the presence of 40 μg/ml of cytosine arabinoside all showed some background activity, probably due to contaminating CEF cellular DNA in the radiolabelled ALVAC DNA probe preparation. However, ALVAC-RG infected CEF cells at 72 hours post-infection exhibited a strong band in the region of approximately 350 kbp representing ALVAC-specific viral DNA accumulation. No such band is detectable when the culture is incubated in the presence of the DNA synthesis inhibitor, cytosine arabinoside. Equivalent samples produced in Vero cells showed a very faint band at approximately 350 kbp in the ALVAC-RG infected Vero cells at time zero. This level represented residual virus. The intensity of the band was amplified at 72 hours post-infection indicating that some level of viral specific DNA replication had occurred in Vero cells which had not resulted in an increase in viral progeny. Equivalent samples produced in MRC-5 cells indicated that no viral specific DNA accumulation was detected under these conditions in this cell line. This experiment was then extended to include additional human cell lines, specifically WISH and Detroit-532 cells. ALVAC infected CEF cells served as a positive control. No viral specific DNA accumulation was detected in either WISH or Detroit cells inoculated with ALVAC-RG. It should be noted that the limits of detection of this method have not been fully ascertained and viral DNA accumulation may be occurring, but at a level below the sensitivity of the method. Other experiments in which viral DNA replication was measured by ³ H-thymidine incorporation support the results obtained with Vero and MRC-5 cells.

Analysis of Rabies Gene Expression. To determine if any viral gene expression, particularly that of the inserted foreign gene, was occurring in the human cell lines even in the absence of viral DNA replication, immunoprecipitation experiments were performed on ³⁵ S-methionine labelled lysates of avian and non-avian cells infected with ALVAC and ALVAC-RG. The results of immunoprecipitation using a rabies G specific monoclonal antibody illustrated specific immunoprecipitation of a 67 kDa glycoprotein in CEF, Vero and MRC-5, WISH and Detroit cells infected with ALVAC-RG. No such specific rabies gene products were detected in any of the uninfected and parentally infected cell lysates.

The results of this experiment indicated that in the human cell lines analyzed, although the ALVAC-RG recombinant was able to initiate an infection and express a foreign gene product under the transcriptional control of the H6 early/late vaccinia virus promoter, the replication did not proceed through DNA replication, nor was there any detectable viral progeny produced. In the Vero cells, although some level of ALVAC-RG specific DNA accumulation was observed, no viral progeny was detected by these methods. These results would indicate that in the human cell lines analyzed the block to viral replication occurs prior to the onset of DNA replication, while in Vero cells, the block occurs following the onset of viral DNA replication.

In order to determine whether the rabies glycoprotein expressed in ALVAC-RG was immunogenic, a number of animal species were tested by inoculation of the recombinant. The efficacy of current rabies vaccines is evaluated in a mouse model system. A similar test was therefore performed using ALVAC-RG. Nine different preparations of virus (including one vaccine batch (J) produced after 10 serial tissue culture passages of the seed virus) with infectious titers ranging from 6.7 to 8.4 log₁₀ TCID₅₀ per ml were serially diluted and 50 to 100 μl of dilutions inoculated into the footpad of four to six week old mice. Mice were challenged 14 days later by the intracranial route with 300 μl of the CVS strain of rabies virus containing from 15 to 43 mouse LD₅₀ as determined by lethality titration in a control group of mice. Potency, expressed as the PD₅₀ (Protective dose 50%), was calculated at 14 days post-challenge. The results of the experiment are shown in Table 6. The results indicated that ALVAC-RG was consistently able to protect mice against rabies virus challenge with a PD₅₀ value ranging from 3.33 to 4.56 with a mean value of 3.73 (STD 0.48). As an extension of this study, male mice were inoculated intracranially with 50 μl of virus containing 6.0 log₁₀ TCID₅₀ of ALVAC-RG or with an equivalent volume of an uninfected cell suspension. Mice were sacrificed on days 1, 3 and 6 post-inoculation and their brains removed, fixed and sectioned. Histopathological examination showed no evidence for neurovirulence of ALVAC-RG in mice.

In order to evaluate the safety and efficacy of ALVAC-RG for dogs and cats, a group of 14, 5 month old beagles and 14, 4 month old cats were analyzed. Four animals in each species were not vaccinated. Five animals received 6.7 log₁₀ TCID₅₀ subcutaneously and five animals received 7.7 log₁₀ TCID₅₀ by the same route. Animals were bled for analysis for anti-rabies antibody. Animals receiving no inoculation or 6.7 log₁₀ TCID₅₀ of ALVAC-RG were challenged at 29 days post-vaccination with 3.7 log₁₀ mouse LD₅₀ (dogs, in the temporal muscle) or 4.3 log₁₀ mouse LD₅₀ (cats, in the neck) of the NYGS rabies virus challenge strain. The results of the experiment are shown in Table 7.

No adverse reactions to inoculation were seen in either cats or dogs with either dose of inoculum virus. Four of 5 dogs immunized with 6.7 log₁₀ TCID₅₀ had antibody titers on day 14 post-vaccination and all dogs had titers at 29 days. All dogs were protected from a challenge which killed three out of four controls. In cats, three of five cats receiving 6.7 log₁₀ TCID₅₀ had specific antibody titers on day 14 and all cats were positive on day 29 although the mean antibody titer was low at 2.9 IU. Three of five cats survived a challenge which killed all controls. All cats immunized with 7.7 log₁₀ TCID₅₀ had antibody titers on day 14 and at day 29 the Geometric Mean Titer was calculated as 8.1 International Units.

The immune response of squirrel monkeys (Saimiri sciureus) to inoculation with ALVAC, ALVAC-RG and an unrelated canarypox virus recombinant was examined. Groups of monkeys were inoculated as described above and sera analyzed for the presence of rabies specific antibody. Apart from minor typical skin reactions to inoculation by the intradermal route, no adverse reactivity was seen in any of the monkeys. Small amounts of residual virus were isolated from skin lesions after intradermal inoculation on days two and four post-inoculation only. All specimens were negative on day seven and later. There was no local reaction to intra-muscular injection. All four monkeys inoculated with ALVAC-RG developed anti-rabies serum neutralizing antibodies as measured in an RFFI test. Approximately six months after the initial inoculation all monkeys and one additional naive monkey were re-inoculated by the subcutaneous route on the external face of the left thigh with 6.5 log₁₀ TCID₅₀ of ALVAC-RG. Sera were analyzed for the presence of anti-rabies antibody. The results are shown in Table 8.

Four of the five monkeys naive to rabies developed a serological response by seven days post-inoculation with ALVAC-RG. All five monkeys had detectable antibody by 11 days post-inoculation. Of the four monkeys with previous exposure to the rabies glycoprotein, all showed a significant increase in serum neutralization titer between days 3 and 7 post-vaccination. The results indicate that vaccination of squirrel monkeys with ALVAC-RG does not produce adverse side-effects and a primary neutralizing antibody response can be induced. An anamnestic response is also induced on re-vaccination. Prior exposure to ALVAC or to a canarypox recombinant expressing an unrelated foreign gene does not interfere with induction of an anti-rabies immune response upon re-vaccination.

The immunological response of HIV-2 seropositive macaques to inoculation with ALVAC-RG was assessed. Animals were inoculated as described above and the presence of anti-rabies serum neutralizing antibody assessed in an RFFI test. The results, shown in Table 9, indicated that HIV-2 positive animals inoculated by the subcutaneous route developed anti-rabies antibody by 11 days after one inoculation. An anamnestic response was detected after a booster inoculation given approximately three months after the first inoculation. No response was detected in animals receiving the recombinant by the oral route. In addition, a series of six animals were inoculated with decreasing doses of ALVAC-RG given by either the intra-muscular or subcutaneous routes. Five of the six animals inoculated responded by 14 days post-vaccination with no significant difference in antibody titer.

Two chimpanzees with prior exposure to HIV were inoculated with 7.0 log₁₀ pfu of ALVAC-RG by the subcutaneous or intra-muscular route. At 3 months post-inoculations both animals were re-vaccinated in an identical fashion. The results are shown in Table 10.

No adverse reactivity to inoculation was noted by either intramuscular or subcutaneous routes. Both chimpanzees responded to primary inoculation by 14 days and a strongly rising response was detected following revaccination.

                  TABLE 1                                                          ______________________________________                                         Sequential Passage of ALVAC in Avian and non-Avian Cells.                                CEF        Vero   MRC-5                                              ______________________________________                                         Pass 1                                                                         Sample   to.sup.a                                                                              2.4          3.0  2.6                                                   t7.sup.b                                                                              7.0          1.4  0.4                                                   t7A.sup.c                                                                             1.2          1.2  0.4                                          Pass 2                                                                         Sample   to     5.0          0.4  .sup. N.D..sup.d                                      t7     7.3          0.4  N.D.                                                  t7A    3.9          N.D. N.D.                                         Pass 3                                                                         Sample   to     5.4          0.4  N.D.                                                  t7     7.4          N.D. N.D.                                                  t7A    3.8          N.D. N.D.                                         Pass 4                                                                         Sample   to     5.2          N.D. N.D.                                                  t7     7.1          N.D. N.D.                                                  t7A    3.9          N.D. N.D.                                         ______________________________________                                          .sup.a This sample was harvested at zero time and represents the residual      input virus. The titer is expressed as log.sub.10 pfu per ml.                  .sup.b This sample was harvested at 7 days postinfection.                      .sup.c This sample was inoculated in the presence of 40 μg/ml of            Cytosine arabinoside and harvested at 7 days post infection.                   .sup.d Not detectable                                                    

                  TABLE 2                                                          ______________________________________                                         Sequential Passage of ALVAC-RG in Avian and non-Avian Cells                             CEF        Vero    MRC-5                                              ______________________________________                                         Pass 1                                                                         Sample  t0.sup.a                                                                              3.0          2.9   2.9                                                  t7.sup.b                                                                              7.1          1.0   1.4                                                  t7A.sup.c                                                                             1.8          1.4   1.2                                          Pass 2                                                                         Sample  t0     5.1          0.4   0.4                                                  t7     7.1          .sup. N.D..sup.d                                                                     N.D.                                                 t7A    3.8          N.D.  N.D.                                         Pass 3                                                                         Sample  t0     5.1          0.4   N.D.                                                 t7     7.2          N.D.  N.D.                                                 t7A    3.6          N.D.  N.D.                                         Pass 4                                                                         Sample  t0     5.1          N.D.  N.D.                                                 t7     7.0          N.D.  N.D.                                                 t7A    4.0          N.D.  N.D                                          ______________________________________                                          .sup.a This sample was harvested at zero time and represents the residual      input virus. The titer is expressed as log.sub.10 pfu per ml.                  .sup.b This sample was harvested at 7 days postinfection.                      .sup.c This sample was inoculated in the presence of 40 μg/ml of            Cytosine arabinoside and harvested at 7 days postinfection.                    .sup.d Not detectable.                                                   

                  TABLE 3                                                          ______________________________________                                         Amplification of residual virus by passage in CEF cells                                CEF        Vero    MRC-5                                               ______________________________________                                         a) ALVAC                                                                       Pass    2.sup.a                                                                              .sup. 7.0.sup.b                                                                             6.0   5.2                                                   3     7.5          4.1   4.9                                                   4     7.5           N.D..sup.c                                                                          N.D.                                                  5     7.1          N.D.  N.D.                                          b) ALVAC-RG                                                                    Pass    2.sup.a                                                                              7.2          5.5   5.5                                                   3     7.2          5.0   5.1                                                   4     7.2          N.D.  N.D.                                                  5     7.2          N.D.  N.D.                                          ______________________________________                                          .sup.a Pass 2 represents the amplification in CEF cells of the 7 day           sample from Pass 1.                                                            .sup.b Titer expressed as log.sub.10 pfu per ml                                .sup.c Not Detectable                                                    

                  TABLE 4                                                          ______________________________________                                         Schedule of inoculation of rhesus macaques with ALVAC-RG (vCP65)               Animal   Inoculation                                                           ______________________________________                                         176L     Primary    1 × 10.sup.8 pfu of vCP65 orally in TANG                      Secondary  1 × 10.sup.7 pfu of vCP65 plus 1 ×                                 10.sup.7                                                                       pfu of vCP82.sup.a by SC route                             185L     Primary    1 × 10.sup.8 pfu of vCP65 orally in TANG                      Secondary  1 × 10.sup.7 pfu of vCP65 plus 1 ×                                 10.sup.7                                                                       pfu of vCP82 by SC route                                   177L     Primary    5 × 10.sup.7 pfu SC of vCP65 by SC route                      Secondary  1 × 10.sup.7 pfu of vCP65 plus 1 ×                                 10.sup.7                                                                       pfu of vCP82 by SC route                                   186L     Primary    5 × 10.sup.7 pfu of vCP65 by SC route                         Secondary  1 × 10.sup.7 pfu of vCP65 plus 1 ×                                 10.sup.7                                                                       pfu of vCP82 by SC route                                   178L     Primary    1 × 10.sup.7 pfu of vCP65 by SC route                182L     Primary    1 × 10.sup.7 pfu of vCP65 by IM route                179L     Primary    1 × 10.sup.6 pfu of vCP65 by SC route                183L     Primary    1 × 10.sup.6 pfu of vCP65 by IM route                180L     Primary    1 × 10.sup.6 pfu of vCP65 by SC route                184L     Primary    1 × 10.sup.5 pfu of vCP65 by IM route                187L     Primary    1 × 10.sup.7 pfu of vCP65 orally                     ______________________________________                                          .sup.a vCP82 is a canarypox virus recombinant expressing the measles viru      fusion and hemagglutinin genes.                                          

                  TABLE 5                                                          ______________________________________                                         Analysis of yield in avian and non-avian cells                                 inoculated with ALVAC-RG                                                       Sample Time                                                                    Cell Type     t0           t72   t72A.sup.b                                    ______________________________________                                         Expt    1                                                                              CEF       3.3.sup.a    7.4 1.7                                                 Vero      3.0          1.4 1.7                                                 MRC-5     3.4          2.0 1.7                                         Expt    2                                                                              CEF       2.9          7.5 <1.7                                                WISH      3.3          2.2 2.0                                                 Detroit-532                                                                              2.8          1.7 <1.7                                        ______________________________________                                          .sup.a Titer expressed as log.sub.10 pfu per ml                                .sup.b Culture incubated in the presence of 40 μg/ml of Cytosine            arabinoside                                                              

                  TABLE 6                                                          ______________________________________                                         Potency of ALVAC-RG as tested in mice                                          Test            Challenge Dose.sup.a                                                                       PD.sub.50.sup.b                                    ______________________________________                                         Initial seed    43          4.56                                               Primary seed    23          3.34                                               Vaccine Batch H 23          4.52                                               Vaccine Batch I 23          3.33                                               Vaccine Batch K 15          3.64                                               Vaccine Batch L 15          4.03                                               Vaccine Batch M 15          3.32                                               Vaccine Batch N 15          3.39                                               Vaccine Batch J 23          3.42                                               ______________________________________                                          .sup.a Expressed as mouse LD.sub.50                                            .sup.b Expressed as log.sub.10 TCID.sub.50                               

                  TABLE 7                                                          ______________________________________                                         Efficacy of ALVAC-RG in dogs and cats                                                  Dogs                 Cats                                              Dose    Antibody.sup.a                                                                          Survival.sup.b                                                                             Antibody                                                                              Survival                                   ______________________________________                                         6.7     11.9     5/5         2.9    3/5                                        7.7     10.1     N.T.        8.1    N.T.                                       ______________________________________                                          .sup.a Antibody at day 29 post inoculation expressed as the geometric mea      titer in International Units.                                                  .sup.b Expressed as a ratio of survivors over animals challenged         

                  TABLE 8                                                          ______________________________________                                         Anti-rabies serological response of Squirrel monkeys                           inoculated with canarypox recombinants                                         Monkey                                                                               Previous Rabies serum-neutralizing antibody.sup.a                        #     Exposure -196.sup.b                                                                             0    3    7    11   21   28                             ______________________________________                                         22    ALVAC.sup.c                                                                             NT.sup.g                                                                               <1.2 <1.2 <1.2 2.1  2.3  2.2                            51    ALVAC.sup.c                                                                             NT      <1.2 <1.2 1.7  2.2  2.2  2.2                            39    vCP37.sup.d                                                                             NT      <1.2 <1.2 1.7  2.1  2.2  N.T..sup.g                     55    vCP37.sup.d                                                                             NT      <1.2 <1.2 1.7  2.2  2.1  N.T.                           37    ALVAC-   2.2     <1.2 <1.2 3.2  3.5  3.5  3.2                                  RG.sup.e                                                                 53    ALVAC-   2.2     <1.2 <1.2 3.6  3.6  3.6  3.4                                  RG.sup.e                                                                 38    ALVAC-   2.7     <1.7 <1.7 3.2  3.8  3.6  N.T.                                 RG.sup.f                                                                 54    ALVAC-   3.2     <1.7 <1.5 3.6  4.2  4.0  3.6                                  RG.sup.f                                                                 57    None     NT      <1.2 <1.2 1.7  2.7  2.7  2.3                            ______________________________________                                          .sup.a As determined by RFFI test on days indicated and expressed in           International Units                                                            .sup.b Day196 represents serum from day 28 after primary vaccination           .sup.c Animals received 5.0 log.sub.10 TCID.sub.50 of ALVAC                    .sup.d Animals received 5.0 log.sub.10 TCID.sub.50 of vCP37                    .sup.e Animals received 5.0 log.sub.10 TCID.sub.50 of ALVACRG                  .sup.f Animals received 7.0 log.sub.10 TCID.sub.50 of ALVACRG                  .sup.g Not tested.                                                       

                                      TABLE 9                                      __________________________________________________________________________     Inoculation of rhesus macaques with ALVAC-RG.sup.a                             Route of Primary Inoculation                                                   Days post-                                                                           or/Tang                                                                               SC SC SC IM SC IM SC IM OR                                        Inoculation                                                                          176L.sup.b                                                                         185L                                                                              177L                                                                              186L                                                                              178L                                                                              182L                                                                              179L                                                                              183L                                                                              180L                                                                              184L                                                                              Y87L.sup.b                                __________________________________________________________________________     -84   --  --                                                                   -9    --  -- -- -- --                                                          3     --  -- -- --                                                             6     --  -- ±                                                                              ±                                                           11    --  -- 16.sup.d                                                                          128                                                            19    --  -- 32 128                                                                               --                                                          35    --  -- 32 512                                                            59    --  -- 64 256                                                            75    --  -- 64 128                                                                               --                                                          99.sup.c                                                                             --  -- 64 256                                                                               -- -- -- -- -- --                                           2     --  -- 32 256                                                                               -- -- -- -- -- -- --                                        6     --  -- 512                                                                               512                                                                               -- -- -- -- -- -- --                                        15    16  16 512                                                                               512                                                                               64 32 64 128                                                                               32 -- --                                        29    16  32 256                                                                               256                                                                               64 64 32 128                                                                               32 -- --                                        55        32          32    32 16 --                                           57    16     128                                                                               128                                                                               16    16          --                                        __________________________________________________________________________      .sup.a See Table 9 for schedule of inoculations.                               .sup.b Animals 176L and 185L received 8.0 log.sub.10 pfu by the oral rout      in 5 ml Tang. Animal 187L received 7.0 log.sub.10 pfu by oral route not i      Tang.                                                                          .sup.c Day of revaccination for animals 176L, 185L, 177L and 186L by S.C.      route, and primary vaccination for animals 178L, 182L, 179L, 183L, 180L,       184L and 187L.                                                                 .sup.d Titers expressed as reciprocal of last dilution showing inhibition      of fluorescence in an RFFI test.                                         

                  TABLE 10                                                         ______________________________________                                         Inoculation of chimpanzees with ALVAC-RG                                       Weeks post-    Animal 431                                                                               Animal 457                                            Inoculation    I.M.      S.C.                                                  ______________________________________                                         0               <8.sup.a <8                                                    1              <8        <8                                                    2               8        32                                                    4              16        32                                                    8              16        32                                                    12.sup.b /0.sup.                                                                              16         8                                                    13/1           128       128                                                   15/3           256       512                                                   20/8           64        128                                                    26/12         32        128                                                   ______________________________________                                          .sup.a Titer expressed as reciprocal of last dilution showing inhibition       of fluorescence in an RFFI test                                                .sup.b Day of reinoculation                                              

Example 9 Immunization of Humans Using Canarypox Expressing Rabies Glycoprotein (ALVAC-RG: vCP65)

ALVAC-RG (vCP65) was generated as described in Example 9 and FIGS. 9A and 9B. For scaling-up and vaccine manufacturing ALVAC-RG (vCP65) was grown in primary CEF derived from specified pathogen free eggs. Cells were infected at a multiplicity of 0.1 and incubated at 37° C. for three days.

The vaccine virus suspension was obtained by ultrasonic disruption in serum free medium of the infected cells; cell debris were then removed by centrifugation and filtration. The resulting clarified suspension was supplemented with lyophilization stabilizer (mixture of amino-acids), dispensed in single dose vials and freeze dried. Three batches of decreasing titer were prepared by ten-fold serial dilutions of the virus suspension in a mixture of serum free medium and lyophilization stabilizer, prior to lyophilization.

Quality control tests were applied to the cell substrates, media and virus seeds and final product with emphasis on the search for adventitious agents and inocuity in laboratory rodents. No undesirable trait was found.

Preclinical data. Studies in vitro indicated that VERO or MRC-5 cells do not support the growth of ALVAC-RG (vCP65); a series of eight (VERO) and 10 (MRC) blind serial passages caused no detectable adaptation of the virus to grow in these non avian lines. Analyses of human cell lines (MRC-5, WISH, Detroit 532, HEL, HNK or EBV-transformed lymphoblastoid cells) infected or inoculated with ALVAC-RG (vCP65) showed no accumulation of virus specific DNA suggesting that in these cells the block in replication occurs prior to DNA synthesis. Significantly, however, the expression of the rabies virus glycoprotein gene in all cell lines tested indicating that the abortive step in the canarypox replication cycle occurs prior to viral DNA replication.

The safety and efficacy of ALVAC-RG (vCP65) were documented in a series of experiments in animals. A number of species including canaries, chickens, ducks, geese, laboratory rodents (suckling and adult mice), hamsters, guinea-pigs, rabbits, cats and dogs, squirrel monkeys, rhesus macaques and chimpanzees, were inoculated with doses ranging from 10⁵ to 10⁸ pfu. A variety of routes were used, most commonly subcutaneous, intramuscular and intradermal but also oral (monkeys and mice) and intracerebral (mice).

In canaries, ALVAC-RG (vCP65) caused a "take" lesion at the site of scarification with no indication of disease or death. Intradermal inoculation of rabbits resulted in a typical poxvirus inoculation reaction which did not spread and healed in seven to ten days. There was no adverse side effects due to canarypox in any of the animal tests. Immunogenicity was documented by the development of anti-rabies antibodies following inoculation of ALVAC-RG (vCP65) in rodents, dogs, cats, and primates, as measured by Rapid Fluorescent Focus Inhibition Test (RFFIT). Protection was also demonstrated by rabies virus challenge experiments in mice, dogs, and cats immunized with ALVAC-RG (vCP65).

Volunteers. Twenty-five healthy adults aged 20-45 with no previous history of rabies immunization were enrolled. Their health status was assessed by complete medical histories, physical examinations, hematological and blood chemistry analyses. Exclusion criteria included pregnancy, allergies, immune depression of any kind, chronic debilitating disease, cancer, injection of immune globins in the past three months, and seropositivity to human immunodeficiency virus (HIV) or to hepatitis B virus surface antigen.

Study design. Participants were randomly allocated to receive either standard Human Diploid Cell Rabies Vaccine (HDC) batch no E0751 (Pasteur Merieux Serums & Vaccine, Lyon, France) or the study vaccine ALVAC-RG (vCP65).

The trial was designated as a dose escalation study. Three batches of experimental ALVAC-RG (vCP65) vaccine were used sequentially in three groups of volunteers (Groups A, B and C) with two week intervals between each step. The concentration of the three batches was 10³.5, 10⁴.5, 10⁵.5 Tissue Culture Infectious Dose (TCID₅₀) per dose, respectively.

Each volunteer received two doses of the same vaccine subcutaneously in the deltoid region at an interval of four weeks. The nature of the injected vaccine was not known by the participants at the time of the first injection but was known by the investigator.

In order to minimize the risk of immediate hypersensitivity at the time of the second injection, the volunteers of Group B allocated to the medium dose of experimental vaccine were injected 1 h previously with the lower dose and those allocated to the higher dose (Group C) received successively the lower and the medium dose at hourly intervals.

Six months later, the recipients of the highest dosage of ALVAC-RG (vCP65) (Group C) and HDC vaccine were offered a third dose of vaccine; they were then randomized to receive either the same vaccine as previously or the alternate vaccine. As a result, four groups were formed corresponding to the following immunization scheme: 1. HDC, HDC-HDC; 2. HDC, HDC-ALVAC-RG (vCP65); 3. ALVAC-RG (vCP65), ALVAC-RG (vCP65)-HDC; 4. ALVAC-RG (vCP65), ALVAC-RG (vCP65), ALVAC-RG (vCP65).

Monitoring of Side Effects. All subjects were monitored for 1 h after injection and re-examined every day for the next five days. They were asked to record local and systemic reactions for the next three weeks and were questioned by telephone two times a week.

Laboratory Investigators. Blood specimens were obtained before enrollment and two, four and six days after each injection. Analysis included complete blood cell count, liver enzymes and creatine kinase assays.

Antibody assays. Antibody assays were performed seven days prior to the first injection and at days 7, 28, 35, 56, 173, 187 and 208 of the study.

The levels of neutralizing antibodies to rabies were determined using the Rapid Fluorescent Focus Inhibition test (RFFIT) (Smith et al., 1973). Canarypox antibodies were measured by direct ELISA. The antigen, a suspension of purified canarypox virus disrupted with 0.1% Triton X100, was coated in microplates. Fixed dilutions of the sera were reacted for two hours at room temperature and reacting antibodies were revealed with a peroxidase labelled antihuman IgG goat serum. The results are expressed as the optical density read at 490 nm.

Analysis. Twenty-five subjects were enrolled and completed the study. There were 10 males and 15 females and the mean age was 31.9 (21 to 48). All but three subjects had evidence of previous smallpox vaccination; the three remaining subjects had no typical scar and vaccination history. Three subjects received each of the lower doses of experimental vaccine (10³.5 and 10⁴.5 TCID₅₀), nine subjects received 10⁵.5 TCID₅₀ and ten received the HDC vaccine.

Safety (Table 11). During the primary series of immunization, fever greater than 37.7° C. was noted within 24 hours after injection in one HDC recipient (37.8° C.) and in one vCP65 10⁵.5 TCID₅₀ recipient (38° C.). No other systemic reaction attributable to vaccination was observed in any participant.

Local reactions were noted in 9/10 recipients of HDC vaccine injected subcutaneously and in 0/3, 1/3 and 9/9 recipients of vCP65 10³.5, 10⁴.5, 10⁵.5 TCID₅₀, respectively.

Tenderness was the most common symptoms and was always mild. Other local symptoms included redness and induration which were also mild and transient. All symptoms usually subsided within 24 hours and never lasted more than 72 hours.

There was no significant change in blood cell counts, liver enzymes or creatine kinase values.

Immune Responses; Neutralizing Antibodies to Rabies (Table 12). Twenty eight days after the first injection all the HDC recipients had protective titers (≧0.5 IU/ml). By contrast none in groups A and B (10³.5 and 10⁴.5 TCID₅₀) and only 2/9 in group C (10⁵.5 TCID₅₀) ALVAC-RG (vCP65) recipients reached this protective titer.

At day 56 (i.e. 28 days after the second injection) protective titers were achieved in 0/3 of Group A, 2/3 of Group B and 9/9 of Group C recipients of ALVAC-RG (vCP65) vaccine and persisted in all 10 HDC recipients.

At day 56 the geometric mean titers were 0.05, 0.47, 4.4 and 11.5 IU/ml in groups A, B, C and HDC respectively.

At day 180, the rabies antibody titers had substantially decreased in all subjects but remained above the minimum protective titer of 0.5 IU/ml in 5/10 HCD recipients and in 5/9 ALVAC-RG (vCP65) recipients; the geometric mean titers were 0.51 and 0.45 IU/ml in groups HCD and C, respectively.

Antibodies to the Canarypox virus (Table 13). The preimmune titers observed varied widely with titers varying from 0.22 to 1.23 O.D. units despite the absence of any previous contact with canary birds in those subjects with the highest titers. When defined as a greater than two-fold increase between preimmunization and post second injection titers, a seroconversion was obtained in 1/3 subjects in group B and in 9/9 subjects in group C whereas no subject seroconverted in groups A or HDC.

Booster Injection. The vaccine was similarly well tolerated six months later, at the time of the booster injection: fever was noted in 2/9 HDC booster recipients and in 1/10 ALVAC-RG (vCP65) booster recipients. Local reactions were present in 5/9 recipients of HDC booster and in 6/10 recipients of the ALVAC-RG (vCP65) booster.

Observations. FIGS. 11A-11D show graphs of rabies neutralizing antibody titers (Rapid Fluorescent Focus Inhibition Test or RFFIT, IU/ml): Booster effect of HDC and vCP65 (10⁵.5 TCID₅₀) in volunteers previously immunized with either the same or the alternate vaccine. Vaccines were given at days 0, 28 and 180. Antibody titers were measured at days 0, 7, 28, 35, 56, 173, and 187 and 208.

As shown in FIGS. 11A to 11D, the booster dose given resulted in a further increase in rabies antibody titers in every subject whatever the immunization scheme. However, the ALVAC-RG (vCP65) booster globally elicited lower immune responses than the HDC booster and the ALVAC-RG (vCP65), ALVAC-RG (vCP65)-ALVAC-RG (vCP65) group had significantly lower titers than the three other groups. Similarly, the ALVAC-RG (vCP65) booster injection resulted in an increase in canarypox antibody titers in 3/5 subjects who had previously received the HDC vaccine and in all five subjects previously immunized with ALVAC-RG (vCP65).

In general, none of the local side effects from administration of vCP65 was indicative of a local replication of the virus. In particular, lesions of the skin such as those observed after injection of vaccine were absent. In spite of the apparent absence of replication of the virus, the injection resulted in the volunteers generating significant amounts of antibodies to both the canarypox vector and to the expressed rabies glycoprotein.

Rabies neutralizing antibodies were assayed with the Rapid Fluorescent Focus Inhibition Test (RFFIT) which is known to correlate well with the sero neutralization test in mice. Of 9 recipients of 10⁵.5 TCID₅₀, five had low level responses after the first dose. Protective titers of rabies antibodies were obtained after the second injection in all recipients of the highest dose tested and even in 2 of the 3 recipients of the medium dose. In this study, both vaccines were given subcutaneously as usually recommended for live vaccines, but not for the inactivated HDC vaccine. This route of injection was selected as it best allowed a careful examination of the injection site, but this could explain the late appearance of antibodies in HDC recipients: indeed, none of the HDC recipients had an antibody increase at day 7, whereas, in most studies where HDC vaccine is give intramuscularly a significant proportion of subjects do (Klietmann et al., Geneva, 1981; Kuwert et al., 1981). However, this invention is not necessarily limited to the subcutaneous route of administration.

The GMT (geometric mean titers) of rabies neutralizing antibodies was lower with the investigational vaccine than with the HDC control vaccine, but still well above the minimum titer required for protection. The clear dose effect response obtained with the three dosages used in this study suggest that a higher dosage might induce a stronger response. Certainly from this disclosure the skilled artisan can select an appropriate dosage for a given patient.

The ability to boost the antibody response is another important result of this Example; indeed, an increase in rabies antibody titers was obtained in every subject after the 6 month dose whatever the immunization scheme, showing that preexisting immunity elicited by either the canarypox vector or the rabies glycoprotein had no blocking effect on the booster with the recombinant vaccine candidate or the conventional HDC rabies vaccine. This contrasts findings of others with vaccinia recombinants in humans that immune response may be blocked by pre-existing immunity (Cooney et al., 1991; Etinger et al., 1991).

Thus, this Example clearly demonstrates that a non-replicating poxvirus can serve as an immunizing vector in humans, with all of the advantages that replicating agents confer on the immune response, but without the safety problem created by a fully permissive virus. And, from this disclosure such as this Example and other Examples suitable dosages and modes or routes for administration or immunization of recombinants containing either rabies or other coding, or expression products thereof, are within the ambit of the skilled artisan as well modes for in vitro expression.

                  TABLE 11                                                         ______________________________________                                         Reactions in the 5 days following vaccination                                  vCP65 dosage                         H D C                                     (TCID50) 10.sup.3.5                                                                               10.sup.4.5                                                                               10.sup.5.5                                                                             control                                   ______________________________________                                         Injection                                                                               1st    2nd    1st  2nd  1st  2nd  1st  2nd                            No. vaccinees                                                                           3      3      3    3    9    9    10   10                             temp >37.7° C.                                                                   0      0      0    0    0    1    1    0                              soreness 0      0      1    1    6    8    8    6                              redness  0      0      0    0    0    4    5    4                              induration                                                                              0      0      0    0    0    4    5    4                              ______________________________________                                    

                  TABLE 12                                                         ______________________________________                                         Rabies neutralizing antibodies (REFIT; IU/ml)                                  Individual titers and geometric mean titers (GMT)                              TCID50/      Days                                                              No.     dose     0        7    28     35   56                                  ______________________________________                                         1       10.sup.3.5                                                                              <0.1     <0.1 <0.1   <0.1 0.2                                 3       10.sup.3.5                                                                              <0.1     <0.1 <0.1   <0.1 <0.1                                4       10.sup.3.5                                                                              <0.1     <0.1 <0.1   <0.1 <0.1                                        G.M.T.   <0.1     <0.1 <0.1   <0.1 <0.1                                6       10.sup.4.5                                                                              <0.1     <0.1 <0.1   <0.1 <0.1                                7       10.sup.4.5                                                                              <0.1     <0.1 <0.1   2.4  1.9                                 10      10.sup.4.5                                                                              <0.1     <0.1 <0.1   1.6  1.1                                         G.M.T.   <0.1     <0.1 0.1    0.58 0.47                                11      10.sup.5.5                                                                              <0.1     <0.1 1.0    3.2  4.3                                 13      10.sup.5.5                                                                              <0.1     <0.1 0.3    6.0  8.8                                 14      10.sup.5.5                                                                              <0.1     <0.1 0.2    2.1  9.4                                 17      10.sup.5.5                                                                              <0.1     <0.1 <0.1   1.2  2.5                                 18      10.sup.5.5                                                                              <0.1     <0.1 0.7    8.3  12.5                                20      10.sup.5.5                                                                              <0.1     <0.1 <0.1   0.3  3.7                                 21      10.sup.5.5                                                                              <0.1     <0.1 0.2    2.6  3.9                                 23      10.sup.5.5                                                                              <0.1     <0.1 <0.1   1.7  4.2                                 25      10.sup.5.5                                                                              <0.1     <0.1 <0.1   0.6  0.9                                         G.M.T.   <0.1     <0.1 0.16   1.9  4.4*                                2       HDC      <0.1     <0.1 0.8    7.1  7.2                                 5       HDC      <0.1     <0.1 9.9    12.8 18.7                                8       HDC      <0.1     <0.1 12.7   21.1 16.5                                9       HDC      <0.1     <0.1 6.0    9.9  14.3                                12      HDC      <0.1     <0.1 5.0    9.2  25.3                                15      HDC      <0.1     <0.1 2.2    5.2  8.6                                 16      HDC      <0.1     <0.1 2.7    7.7  20.7                                19      HDC      <0.1     <0.1 2.6    9.9  9.1                                 22      HDC      <0.1     <0.1 1.4    8.6  6.6                                 24      HDC      <0.1     <0.1 0.8    5.8  4.7                                         G.M.T.   <0.1     <0.1 2.96   9.0  11.5*                               ______________________________________                                          p = 0.007 student t test                                                 

                  TABLE 13                                                         ______________________________________                                         Canarypox antibodies: ELISA Geometric Mean Titers*                             VCP65                                                                          dosage     Days                                                                TCID50/dose                                                                               0         7      28     35   56                                     ______________________________________                                         10.sup.3.5 0.69      ND     0.76   ND   0.68                                   10.sup.4.5 0.49      0.45   0.56   0.63 0.87                                   10.sup.5.5 0.38      0.38   0.77   1.42 1.63                                   HDC control                                                                               0.45      0.39   0.40   0.35 0.39                                   ______________________________________                                          *optical density at 1/25 dilution                                        

Example 10 Comparison of the LD₅₀ of ALVAC and NYVAC with Various Vaccinia Virus Strains

Mice. Male outbred Swiss Webster mice were purchased from Taconic Farms (Germantown, N.Y.) and maintained on mouse chow and water ad libitum until use at 3 weeks of age ("normal" mice). Newborn outbred Swiss Webster mice were of both sexes and were obtained following timed pregnancies performed by Taconic Farms. All newborn mice used were delivered within a two day period.

Viruses. ALVAC was derived by plaque purification of a canarypox virus population and was prepared in primary chick embryo fibroblast cells (CEF). Following purification by centrifugation over sucrose density gradients, ALVAC was enumerated for plaque forming units in CEF cells. The WR(L) variant of vaccinia virus was derived by selection of large plaque phenotypes of WR (Panicali et al., 1981). The Wyeth New York State Board of Health vaccine strain of vaccinia virus was obtained from Pharmaceuticals Calf Lymph Type vaccine Dryvax, control number 302001B. Copenhagen strain vaccinia virus VC-2 was obtained from Institut Merieux, France. Vaccinia virus strain NYVAC was derived from Copenhagen VC-2. All vaccinia virus strains except the Wyeth strain were cultivated in Vero African green monkey kidney cells, purified by sucrose gradient density centrifugation and enumerated for plaque forming units on Vero cells. The Wyeth strain was grown in CEF cells and enumerated for plaque forming units in CEF cells.

Inoculations. Groups of 10 normal mice were inoculated intracranially (ic) with 0.05 ml of one of several dilutions of virus prepared by 10-fold serially diluting the stock preparations in sterile phosphate-buffered saline. In some instances, undiluted stock virus preparation was used for inoculation.

Groups of 10 newborn mice, 1 to 2 days old, were inoculated ic similarly to the normal mice except that an injection volume of 0.03 ml was used.

All mice were observed daily for mortality for a period of 14 days (newborn mice) or 21 days (normal mice) after inoculation. Mice found dead the morning following inoculation were excluded due to potential death by trauma.

The lethal dose required to produce mortality for 50% of the experimental population (LD₅₀) was determined by the proportional method of Reed and Muench (Reed and Muench, 1938).

Comparison of the LD₅₀ of ALVAC and NYVAC with Various Vaccinia Virus Strains for Normal, Young Outbred Mice by the ic Route. In young, normal mice, the virulence of NYVAC and ALVAC were several orders of magnitude lower than the other vaccinia virus strains tested (Table 14). NYVAC and ALVAC were found to be over 3,000 times less virulent in normal mice than the Wyeth strain; over 12,500 times less virulent than the parental VC-2 strain; and over 63,000,000 times less virulent than the WR(L) variant. These results would suggest that NYVAC is highly attenuated compared to other vaccinia strains, and that ALVAC is generally nonvirulent for young mice when administered intracranially, although both may cause mortality in mice at extremely high doses (3.85×10⁸ PFUs, ALVAC and 3×10⁸ PFUs, NYVAC) by an undetermined mechanism by this route of inoculation.

Comparison of the LD₅₀ of ALVAC and NYVAC with Various Vaccinia Virus Strains for Newborn Outbred Mice by the ic Route. The relative virulence of 5 poxvirus strains for normal, newborn mice was tested by titration in an intracranial (ic) challenge model system (Table 15). With mortality as the endpoint, LD₅₀ values indicated that ALVAC is over 100,000 times less virulent than the Wyeth vaccine strain of vaccinia virus; over 200,000 times less virulent than the Copenhagen VC-2 strain of vaccinia virus; and over 25,000,000 times less virulent than the WR-L variant of vaccinia virus. Nonetheless, at the highest dose tested, 6.3×10⁷ PFUs, 100% mortality resulted. Mortality rates of 33.3% were observed at 6.3×10⁶ PFUs. The cause of death, while not actually determined, was not likely of toxicological or traumatic nature since the mean survival time (MST) of mice of the highest dosage group (approximately 6.3 LD₅₀) was 6.7±1.5 days. When compared to WR(L) at a challenge dose of 5 LD₅₀, wherein MST is 4.8±0.6 days, the MST of ALVAC challenged mice was significantly longer (P=0.001).

Relative to NYVAC, Wyeth was found to be over 15,000 times more virulent; VC-2, greater than 35,000 times more virulent; and WR(L), over 3,000,000 times more virulent. Similar to ALVAC, the two highest doses of NYVAC, 6×10⁸ and 6×10⁷ PFUs, caused 100% mortality. However, the MST of mice challenged with the highest dose, corresponding to 380 LD₅₀, was only 2 days (9 deaths on day 2 and 1 on day 4). In contrast, all mice challenged with the highest dose of WR-L, equivalent to 500 LD₅₀, survived to day 4.

                  TABLE 14                                                         ______________________________________                                         Calculated 50% Lethal                                                          Dose for mice by                                                               various vaccinia                                                               virus strains and for                                                          canarypox virus                                                                (ALVAC) by the ic                                                              route.                                                                         POXVIRUS           CALCULATED                                                  STRAIN             LD.sub.50 (PFUs)                                            ______________________________________                                         WR(L)              2.5                                                         VC-2               1.26 × 10.sup.4                                       WYETH              5.00 × 10.sup.4                                       NYVAC              1.58 × 10.sup.8                                       ALVAC              1.58 × 10.sup.8                                       ______________________________________                                    

                  TABLE 15                                                         ______________________________________                                         Calculated 50% Lethal                                                          Dose for newborn mice                                                          by various vaccinia                                                            virus strains and for                                                          canarypox virus                                                                (ALVAC) by the ic                                                              route.                                                                         POXVIRUS           CALCULATED                                                  STRAIN             LD.sub.50 (PFUs)                                            ______________________________________                                         WR(L)              0.4                                                         VC-2               0.1                                                         WYETH              1.6                                                         NYVAC              1.58 × 10.sup.6                                       ALVAC              1.00 × 10.sup.7                                       ______________________________________                                    

Example 11 Evaluation of NYVAC (vP866) and NYVAC-RG (vP879)

Immunoprecipitations. Preformed monolayers of avian or non-avian cells were inoculated with 10 pfu per cell of parental NYVAC (vP866) or NYVAC-RG (vP879) virus. The inoculation was performed in EMEM free of methionine and supplemented with 2% dialyzed fetal bovine serum. After a one hour incubation, the inoculum was removed and the medium replaced with EMEM (methionine free) containing 20 μCi/ml of ³⁵ S-methionine. After an overnight incubation of approximately 16 hours, cells were lysed by the addition of Buffer A (1% Nonidet P-40, 10 mM Tris pH7.4, 150 mM NaCl, 1 mM EDTA, 0.01% sodium azide, 500 units per ml of aprotinin, and 0.02% phenyl methyl sulfonyl fluoride). Immunoprecipitation was performed using a rabies glycoprotein specific monoclonal antibody designated 24-3F10 supplied by Dr. C. Trinarchi, Griffith Laboratories, New York State Department of Health, Albany, N.Y., and a rat anti-mouse conjugate obtained from Boehringer Mannheim Corporation (Cat. #605-500). Protein A Sepharose CL-48 obtained from Pharmacia LKB Biotechnology Inc., Piscataway, N.J., was used as a support matrix. Immunoprecipitates were fractionated on 10% polyacrylamide gels according to the method of Dreyfuss et. al. (1984). Gels were fixed, treated for fluorography with 1M Na-salicylate for one hour, and exposed to Kodak XAR-2 film to visualize the immunoprecipitated protein species.

Sources of Animals. New Zealand White rabbits were obtained from Hare-Marland (Hewitt, N.J.). Three week old male Swiss Webster outbred mice, timed pregnant female Swiss Webster outbred mice, and four week old Swiss Webster nude (nu⁺ nu⁺) mice were obtained from Taconic Farms, Inc. (Germantown, N.Y.). All animals were maintained according to NIH guidelines. All animal protocols were approved by the institutional IACUC. When deemed necessary, mice which were obviously terminally ill were euthanized.

Evaluation of Lesions in Rabbits. Each of two rabbits was inoculated intradermally at multiple sites with 0.1 ml of PBS containing 10⁴, 10⁵, 10⁶, 10⁷, or 10⁸ pfu of each test virus or with PBS alone. The rabbits were observed daily from day 4 until lesion resolution. Indurations and ulcerations were measured and recorded.

Virus Recovery from Inoculation Sites. A single rabbit was inoculated intradermally at multiple sites with 0.1 ml of PBS containing 10⁶, 10⁷, or 10⁸ pfu of each test virus or with PBS alone. After 11 days, the rabbit was euthanized and skin biopsy specimens taken from each of the inoculation sites were aseptically prepared by mechanical disruption and indirect sonication for virus recovery. Infectious virus was assayed by plaque titration on CEF monolayers.

Virulence in Mice. Groups of ten mice, or five in the nude mice experiment, were inoculated ip with one of several dilutions of virus in 0.5 ml of sterile PBS. Reference is also made to Example 11.

Cyclophosphamide (CY) Treatment. Mice were injected by the ip route with 4 mg (0.02 ml) of CY (SIGMA) on day -2, followed by virus injection on day 0. On the following days post infection, mice were injected ip with CY: 4 mg on day 1; 2 mg on days 4, 7 and 11; 3 mg on days 14, 18, 21, 25 and 28. Immunosuppression was indirectly monitored by enumerating white blood cells with a Coulter Counter on day 11. The average white blood cell count was 13,500 cells per μl for untreated mice (n=4) and 4,220cells per μl for CY-treated control mice (n=5).

Calculation of LD₅₀. The lethal dose required to produce 50% mortality (LD₅₀) was determined by the proportional method of Reed and Muench (Reed and Muench 1938).

Potency Testing of NYVAC-RG in Mice. Four to six week old mice were inoculated in the footpad with 50 to 100 μl of a range of dilutions (2.0-8.0 log₁₀ tissue culture infective dose 50% (TCID₅₀)) of either VV-RG (Kieny et al., 1984), ALVAC-RG (Taylor et al., 1991b), or the NYVAC-RG. Each group consisted of eight mice. At 14 days post-vaccination, the mice were challenged by intracranial inoculation with 15 LD₅₀ of the rabies virus CVS strain (0.03 ml). On day 28, surviving mice were counted and protective does 50% (PD₅₀) calculated.

Derivation of NYVAC (vP866). The NYVAC strain of vaccinia virus was generated from VC-2, a plaque cloned isolate of the COPENHAGEN vaccine strain. To generate NYVAC from VC-2, eighteen vaccinia ORFs, including a number of viral functions associated with virulence, were precisely deleted in a series of sequential manipulations as described earlier in this disclosure. These deletions were constructed in a manner designed to prevent the appearance of novel unwanted open reading frames. FIG. 10 schematically depicts the ORFs deleted to generate NYVAC. At the top of FIG. 10 is depicted the HindIII restriction map of the vaccinia virus genome (VC-2 plaque isolate, COPENHAGEN strain). Expanded are the six regions of VC-2 that were sequentially deleted in the generation of NYVAC. The deletions were described earlier in this disclosure (Examples 1 through 6). Below such deletion locus is listed the ORFs which were deleted from that locus, along with the functions or homologies and molecular weight of their gene products.

Replication Studies of NYVAC and ALVAC on Human Tissue Cell Lines. In order to determine the level of replication of NYVAC strain of vaccinia virus (vP866) in cells of human origin, six cell lines were inoculated at an input multiplicity of 0.1 pfu per cell under liquid culture and incubated for 72 hours. The COPENHAGEN parental clone (VC-2) was inoculated in parallel. Primary chick embryo fibroblast (CEF) cells (obtained from 10-11 day old embryonated eggs of SPF origin, Spafas, Inc., Storrs, Conn.) were included to represent a permissive cell substrate for all viruses. Cultures were analyzed on the basis of two criteria: the occurrence of productive viral replication and expression of an extrinsic antigen.

The replication potential of NYVAC in a number of human derived cells are shown in Table 16. Both VC-2 and NYVAC are capable of productive replication in CEF cells, although NYVAC with slightly reduced yields. VC-2 is also capable of productive replication in the six human derived cell lines tested with comparable yields except in the EBV transformed lymphoblastoid cell line JT-1 (human lymphoblastoid cell line transformed with Epstein-Barr virus, see Rickinson et al., 1984). In contrast, NYVAC is highly attenuated in its ability to productively replicate in any of the human derived cell lines tested. Small increases of infectious virus above residual virus levels were obtained from NYVAC-infected MRC-5 (ATCC #CCL171, human embryonic lung origin), DETROIT 532 (ATCC #CCL54, human foreskin, Downs Syndrome), HEL 299 (ATCC #CCL137, human embryonic lung cells) and HNK (human neonatal kidney cells, Whittiker Bioproducts, Inc. Walkersville, Md., Cat #70-151) cells. Replication on these cell lines was significantly reduced when compared to virus yields obtained from NYVAC-infected CEF cells or with parental VC-2 (Table 16). It should be noted that the yields at 24 hours in CEF cells for both NYVAC and VC-2 is equivalent to the 72-hour yield. Allowing the human cell line cultures to incubate an additional 48 hours (another two viral growth cycles) may, therefore, have amplified the relative virus yield obtained.

Consistent with the low levels of virus yields obtained in the human-derived cell lines, MRC-5 and DETROIT 532, detectable but reduced levels of NYVAC-specific DNA accumulation were noted. The level of DNA accumulation in the MRC-5 and DETROIT 532 NYVAC-infected cell lines relative to that observed in NYVAC-infected CEF cells paralleled the relative virus yields. NYVAC-specific viral DNA accumulation was not observed in any of the other human-derived cells.

An equivalent experiment was also performed using the avipox virus, ALVAC. The results of virus replication are also shown in Table 16. No progeny virus was detectable in any of the human cell lines consistent with the host range restriction of canarypox virus to avian species. Also consistent with a lack of productive replication of ALVAC in these human-derived cells is the observation that no ALVAC-specific DNA accumulation was detectable in any of the human-derived cell lines.

Expression of Rabies Glycoprotein by NYVAC-RG (vP879) in Human Cells. In order to determine whether efficient expression of a foreign gene could be obtained in the absence of significant levels of productive viral replication, the same cell lines were inoculated with the NYVAC recombinant expressing the rabies virus glycoprotein (vP879, Example 7) in the presence of ³⁵ S-methionine. Immunoprecipitation of the rabies glycoprotein was performed from the radiolabelled culture lysate using a monoclonal antibody specific for the rabies glycoprotein. Immunoprecipitation of a 67 kDa protein was detected consistent with a fully glycosylated form of the rabies glycoprotein. No serologically crossreactive product was detected in uninfected or parental NYVAC infected cell lysates. Equivalent results were obtained with all other human cells analyzed.

Inoculations on the Rabbit Skin. The induction and nature of skin lesions on rabbits following intradermal (id) inoculations has been previously used as a measure of pathogenicity of vaccinia virus strains (Buller et al., 1988; Child et al., 1990; Fenner, 1958, Flexner et al., 1987; Ghendon and Chernos 1964). Therefore, the nature of lesions associated with id inoculations with the vaccinia strains WR (ATCC #VR119 plaque purified on CV-1 cells, ATCC #CCL70, and a plaque isolate designated L variant, ATCC #VR2035 selected, as described in Panicali et al., 1981)), WYETH (ATCC #VR325 marketed as DRYVAC by Wyeth Laboratories, Marietta, Pa.), COPENHAGEN (VC-2), and NYVAC was evaluated by inoculation of two rabbits (A069 and A128). The two rabbits displayed different overall sensitivities to the viruses, with rabbit A128 displaying less severe reactions than rabbit A069. In rabbit A128, lesions were relatively small and resolved by 27 days post-inoculation. On rabbit A069, lesions were intense, especially for the WR inoculation sites, and resolved only after 49 days. Intensity of the lesions was also dependent on the location of the inoculation sites relative to the lymph drainage network. In particular, all sites located above the backspine displayed more intense lesions and required longer times to resolve the lesions located on the flanks. All lesions were measured daily from day 4 to the disappearance of the last lesion, and the means of maximum lesion size and days to resolution were calculated (Table 17). No local reactions were observed from sites injected with the control PBS. Ulcerative lesions were observed at sites injected with WR, VC-2 and WYETH vaccinia virus strains. Significantly, no induration or ulcerative lesions were observed at sites of inoculation with NYVAC.

Persistence of Infectious Virus at the Site of Inoculation. To assess the relative persistence of these viruses at the site of inoculation, a rabbit was inoculated intradermally at multiple sites with 0.1 ml PBS containing 10⁶, 10⁷ or 10⁸ pfu of VC-2, WR, WYETH or NYVAC. For each virus, the 10⁷ pfu dose was located above the backspine, flanked by the 10⁶ and 10⁸ doses. Sites of inoculation were observed daily for 11 days. WR elicited the most intense response, followed by VC-2 and WYETH (Table 18). Ulceration was first observed at day 9 for WR and WYETH and day 10 for VC-2. Sites inoculated with NYVAC or control PBS displayed no induration or ulceration. At day 11 after inoculation, skin samples from the sites of inoculation were excised, mechanically disrupted, and virus was titrated on CEF cells. The results are shown in Table 18. In no case was more virus recovered at this timepoint than was administered. Recovery of vaccinia strain, WR, was approximately 10⁶ pfu of virus at each site irrespective of amount of virus administered. Recovery of vaccinia strains WYETH and VC-2 was 10³ to 10⁴ pfu regardless of amount administered. No infectious virus was recovered from sites inoculated with NYVAC.

Inoculation of Genetically or Chemically Immune Deficient Mice. Intraperitoneal inoculation of high doses of NYVAC (5×10⁸ pfu) or ALVAC (10⁹ pfu) into nude mice caused no deaths, no lesions, and no apparent disease through the 100 day observation period. In contrast, mice inoculated with WR (10³ to 10⁴ pfu), WYETH (5×10⁷ or 5×10⁸ pfu) or VC-2 (10⁴ to 10⁹ pfu) displayed disseminated lesions typical of poxviruses first on the toes, then on the tail, followed by severe orchitis in some animals. In mice infected with WR or WYETH, the appearance of disseminated lesions generally led to eventual death, whereas most mice infected with VC-2 eventually recovered. Calculated LD₅₀ values are given in Table 19.

In particular, mice inoculated with VC-2 began to display lesions on their toes (red papules) and 1 to 2 days later on the tail. These lesions occurred between 11 and 13 days post-inoculation (pi) in mice given the highest doses (10⁹, 10⁸, 10⁷ and 10⁶ pfu), on day 16 pi in mice given 10⁵ pfu and on day 21 pi in mice given 10⁴ pfu. No lesions were observed in mice inoculated with 10³ and 10² pfu during the 100 day observation period. Orchitis was noticed on day 23 pi in mice given 10⁹ and 10⁸ pfu, and approximately 7 days later in the other groups (10⁷ to 10⁴ pfu). Orchitis was especially intense in the 10⁹ and 10⁸ pfu groups and, although receding, was observed until the end of the 100 day observation period. Some pox-like lesions were noticed on the skin of a few mice, occurring around 30-35 days pi. Most pox lesions healed normally between 60-90 days pi. only one mouse died in the group inoculated with 10⁹ pfu (Day 34 pi) and one mouse died in the group inoculated with 10⁸ pfu (Day 94 pi). No other deaths were observed in the VC-2 inoculated mice.

Mice inoculated with 10⁴ pfu of the WR strain of vaccinia started to display pox lesions on Day 17 pi. These lesions appeared identical to the lesions displayed by the VC-2 injected mice (swollen toes, tail). Mice inoculated with 10³ pfu of the WR strain did not develop lesions until 34 days pi. Orchitis was noticed only in the mice inoculated with the highest dose of WR (10⁴ pfu). During the latter stages of the observation period, lesions appeared around the mouth and the mice stopped eating. All mice inoculated with 10⁴ pfu of WR died or were euthanized when deemed necessary between 21 days and 31 days pi. Four out of the 5 mice injected with 10³ pfu of WR died or were euthanized when deemed necessary between 35 days and 57 days pi. No deaths were observed in mice inoculated with lower doses of WR (1 to 100 pfu).

Mice inoculated with the WYETH strain of vaccinia virus at higher doses 5×10⁷ and 5×10⁸ pfu) showed lesions on toes and tails, developed orchitis, and died. Mice injected with 5×10⁶ pfu or less of WYETH showed no signs of disease or lesions.

As shown in Table 19, CY-treated mice provided a more sensitive model for assaying poxvirus virulence than did nude mice. LD₅₀ values for the WR, WYETH, and VC-2 vaccinia virus strains were significantly lower in this model system than in the nude mouse model. Additionally, lesions developed in mice injected with WYETH, WR and VC-2 vaccinia viruses, as noted below, with higher doses of each virus resulting in more rapid formation of lesions. As was seen with nude mice, CY-treated mice injected with NYVAC or ALVAC did not develop lesions. However, unlike nude mice, some deaths were observed in CY-treated mice challenged with NYVAC or ALVAC, regardless of the dose. These random incidences are suspect as to the cause of death.

Mice injected with all doses of WYETH (9.5×10⁴ to 9.5×10⁸ pfu) displayed pox lesions on their tail and/or on their toes between 7 and 15 days pi. In addition, the tails and toes were swollen. Evolution of lesions on the tail was typical of pox lesions with formation of a papule, ulceration and finally formation of a scab. Mice inoculated with all doses of VC-2 (1.65×10⁵ to 1.65×10⁹) also developed pox lesions on their tails and/or their toes analogous to those of WYETH injected mice. These lesions were observed between 7-12 days post inoculation. No lesions were observed on mice injected with lower doses of WR virus, although deaths occurred in these groups.

Potency Testing of NYVAC-RG. In order to determine that attenuation of the COPENHAGEN strain of vaccinia virus had been effected without significantly altering the ability of the resulting NYVAC strain to be a useful vector, comparative potency tests were performed. In order to monitor the immunogenic potential of the vector during the sequential genetic manipulations performed to attenuate the virus, a rabiesvirus glycoprotein was used as a reporter extrinsic antigen. The protective efficacy of the vectors expressing the rabies. glycoprotein gene was evaluated in the standard NIH mouse potency test for rabies (Seligmann, 1973). Table 20 demonstrates that the PD₅₀ values obtained with the highly attenuated NYVAC vector are identical to those obtained using a COPENHAGEN-based recombinant containing the rabies glycoprotein gene in the tk locus (Kieny et al., 1984) and similar to PD₅₀ values obtained with ALVAC-RG, a canarypox based vector restricted to replication to avian species.

Observations. NYVAC, deleted of known virulence genes and having restricted in vitro growth characteristics, was analyzed in animal model systems to assess its attenuation characteristics. These studies were performed in comparison with the neurovirulent vaccinia virus laboratory strain, WR, two vaccinia virus vaccine strains, WYETH (New York City Board of Health) and COPENHAGEN (VC-2), as well as with a canarypox virus strain, ALVAC (See also Example 11). Together, these viruses provided a spectrum of relative pathogenic potentials in the mouse challenge model and the rabbit skin model, with WR being the most virulent strain, WYETH and COPENHAGEN (VC-2) providing previously utilized attenuated vaccine strains with documented characteristics, and ALVAC providing an example of a poxvirus whose replication is restricted to avian species. Results from these in vivo analyses clearly demonstrate the highly attenuated properties of NYVAC relative to the vaccinia virus strains, WR, WYETH and COPENHAGEN (VC-2) (Tables 14-20). Significantly, the LD₅₀ values for NYVAC were comparable to those observed with the avian host restricted avipoxvirus, ALVAC. Deaths due to NYVAC, as well as ALVAC, were observed only when extremely high doses of virus were administered via the intracranial route (Example 11, Tables 14, 15, 19). It has not yet been established whether these deaths were due to nonspecific consequences of inoculation of a high protein mass. Results from analyses in immunocompromised mouse models (nude and CY-treated) also demonstrate the relatively high attenuation characteristics of NYVAC, as compared to WR, WYETH and COPENHAGEN strains (Tables 17 and 18). Significantly, no evidence of disseminated vaccinia infection or vaccinial disease was observed in NYVAC-inoculated animals or ALVAC-inoculated animals over the observation period. The deletion of multiple virulence-associated genes in NYVAC shows a synergistic effect with respect to pathogenicity. Another measure of the inocuity of NYVAC was provided by the intradermal administration on rabbit skin (Tables 17 and 18). Considering the results with ALVAC, a virus unable to replicate in nonavian species, the ability to replicate at the site of inoculation is not the sole correlate with reactivity, since intradermal inoculation of ALVAC caused areas of induration in a dose dependent manner. Therefore, it is likely that factors other than the replicative capacity of the virus contribute to the formation of the lesions. Deletion of specific virulence-associated genes in NYVAC prevents lesion occurrence.

Together, the results in this Example and in foregoing Examples, including Example 10, demonstrate the highly attenuated nature of NYVAC relative to WR, and the previously utilized vaccinia virus vaccine strains, WYETH and COPENHAGEN. In fact, the pathogenic profile of NYVAC, in the animal model systems tested, was similar to that of ALVAC, a poxvirus known to productively replicate only in avian species. The apparently restricted capacity of NYVAC to productively replicate on cells derived from humans (Table 16) and other species, including the mouse, swine, dog and horse, provides a considerable barrier that limits or prevents potential transmission to unvaccinated contacts or to the general environment in addition to providing a vector with reduced probability of dissemination within the vaccinated individual.

Significantly, NYVAC-based vaccine candidates have been shown to be efficacious. NYVAC recombinants expressing foreign gene products from a number of pathogens have elicited immunological responses towards the foreign gene products in several animal species, including primates. In particular, a NYVAC-based recombinant expressing the rabies glycoprotein was able to protect mice against a lethal rabies challenge. The potency of the NYVAC-based rabies glycoprotein recombinant was comparable to the PD₅₀ value for a COPENHAGEN-based recombinant containing the rabies glycoprotein in the tk locus (Table 20). NYVAC-based recombinants have also been shown to elicit measles virus neutralizing antibodies in rabbits and protection against pseudorabies virus and Japanese encephalitis virus challenge in swine. The highly attenuated NYVAC strain confers safety advantages with human, animal, medical and veterinary applications (Tartaglia et al., 1992). Furthermore, the use of NYVAC as a general laboratory expression vector system may greatly reduce the biological hazards associated with using vaccinia virus.

By the following criteria, the results of this Example and the Examples herein, including Example 10, show NYVAC to be highly attenuated: a) no detectable induration or ulceration at site of inoculation (rabbit skin); b) rapid clearance of infectious virus from intradermal site of inoculation (rabbit. skin); c) absence of testicular inflammation (nude mice); d) greatly reduced virulence (intracranial challenge, both three-week old and newborn mice); e) greatly reduced pathogenicity and failure to disseminate in immunodeficient subjects (nude and cyclophosphamide treated mice); and f) dramatically reduced ability to replicate on a variety of human tissue culture cells. Yet, in spite of being highly attenuated, NYVAC, as a vector, retains the ability to induce strong immune responses to extrinsic antigens.

                  TABLE 16                                                         ______________________________________                                         Replication of COPENHAGEN (VC-2), NYVAC and                                    ALVAC in avian or human derived cell lines                                             Hours                                                                          post-     Yield.sup.a         %                                        Cells   infection VC-2    NYVAC   ALVAC   Yield                                ______________________________________                                         CEF     0         3.8.sup.b                                                                              3.7     4.5                                                  24        8.3     7.8     6.6                                                  48        8.6     7.9     77                                                   72        8.3     7.7     7.5     25                                           72A.sup.c <1.4    1.8     3.1                                          MRC-5   0         3.8     3.8     4.7                                                  72        7.2     4.6     3.8     0.25                                         72A       2.2     2.2     3.7                                          WISH*   0         3.4     3.4     4.3                                                  72        7.6     2.2     3.1     0.000                                                                          4                                            72A       --.sup.d                                                                               1.9     2.9                                          DETROIT 0         3.8     3.7     4.4                                                  72        7.2     5.4     3.4     1.6                                          72A       1.7     1.7     2.9                                          HEL     0         3.8     3.5     4.3                                                  72        7.5     4.6     3.3     0.125                                        72A       2.5     2.1     3.6                                          JT-1    0         3.1     3.1     4.1                                                  72        6.5     3.1     4.2     0.039                                        72A       2.4     2.1     4.4                                          HNK     0         3.8     3.7     4.7                                                  72        7.6     4.5     3.6     0.079                                        72A       3.1     2.7     3.7                                          ______________________________________                                          .sup.a Yield of NYVAC at 72 hours postinfection expressed as a percentage      of yield of VAC2 after 72 hours on the same cell line.                         .sup.b Titer expressed as LOG.sub.50 pfu per ml.                               .sup.c Sample was incubated in the presence of 40 μg/ml of cytosine         arabinoside.                                                                   .sup.d Not determined.                                                         *ATCC #CCL25 Human amnionic cells.                                       

                  TABLE 17                                                         ______________________________________                                         Induration and ulceration at the site of                                       intradermal inoculation of the rabbit skin                                     VIRUS               INDURATION   ULCERATION                                    STRAIN   DOSE.sup.a Size.sup.b                                                                            Days.sup.c                                                                             Size Days                                   ______________________________________                                         WR       10.sup.4   386    30      88   30                                              10.sup.5   622    35      149  32                                              10.sup.6   1057   34      271  34                                              10.sup.7   877    35      204  35                                              10.sup.8   581    25      88   26                                     WYETH    10.sup.4   32     5       --.sup.d                                                                            --                                              10.sup.5   116    15      --   --                                              10.sup.6   267    17      3    15                                              10.sup.7   202    17      3    24                                              10.sup.8   240    29      12   31                                     VC-2     10.sup.4   64     7       --   --                                              10.sup.5   86     8       --   --                                              10.sup.6   136    17      --   --                                              10.sup.7   167    21      6    10                                              10.sup.8   155    32      6    8                                      NYVAC    10.sup.4   --     --      --   --                                              10.sup.5   --     --      --   --                                              10.sup.6   --     --      --   --                                              10.sup.7   --     --      --   --                                              10.sup.8   --     --      --   --                                     ______________________________________                                          .sup.a pfu of indicated vaccinia virus in 0.1 ml PBS inoculated                intradermally into one site.                                                   .sup.b mean maximum size of lesions (mm.sup.2)                                 .sup.c mean time after inoculation for complete healing of lesion.             .sup.d no lesions discernable.                                           

                  TABLE 18                                                         ______________________________________                                         Persistence of poxviruses at the site of                                       intradermal inoculation                                                                                 Total Virus                                           Virus         Inoculum Dose                                                                             Recovered                                             ______________________________________                                         WR            8.0.sup.a  6.14                                                                7.0        6.26                                                                6.0        6.21                                                  WYETH         8.0        3.66                                                                7.0        4.10                                                                6.0        3.59                                                  VC-2          8.0        4.47                                                                7.0        4.74                                                                6.0        3.97                                                  NYVAC         8.0        0                                                                   7.0        0                                                                   6.0        0                                                     ______________________________________                                          .sup.a expressed as log.sub.10 pfu.                                      

                  TABLE 19                                                         ______________________________________                                         Virulence studies in immunocompromised mice                                              LD.sub.50.sup.a                                                      Poxvirus                 Cyclophosphamide                                      Strain      Nude mice    treated mice                                          ______________________________________                                         WR          422          42                                                    VC-2        >10.sup.9    <1.65 × 10.sup.5                                WYETH       1.58 × 10.sup.7                                                                       1.83 × 10.sup.6                                 NYVAC       >5.50 × 10.sup.8                                                                      7.23 × 108                                      ALVAC       >10.sup.9    ≧5.00 × 10.sup.8b                        ______________________________________                                          .sup.a Calculated 50% lethal dose (pfu) for nude or cyclophosphamide           treated mice by the indicated vaccinia viruses and for ALVAC by                intraperitoneal route.                                                         .sup.b 5 out of 10 mice died at the highest dose of 5 × 10.sup.8         pfu.                                                                     

                  TABLE 20                                                         ______________________________________                                         Comparative efficacy of NYVAC-RG and ALVAC-RG                                  in mice                                                                               Recombinant                                                                            PD.sub.50.sup.a                                                 ______________________________________                                                VV-RG   3.74                                                                   ALVAC-RG                                                                               3.86                                                                   NYVAC-RG                                                                               3.70                                                            ______________________________________                                          .sup.a Four to six week old mice were inoculated in the footpad with 5010      μl of a range of dilutions (2.0-8.0 log.sub.10 tissue culture infectio      dose 50% (TCID.sub.50) of either the VVRG (Kieny et al., 1984), ALVACRG        (vCP65) or NYVACRG (vP879). At day 14, mice of each group were challenged      by intracranial inoculation of 30 μl of a live CVS strain rabies virus      corresponding to 15 lethal dose 50% (LD.sub.50) per mouse. At day 28,          surviving  #mice were counted and a protective dose 50% (PD.sub.5) was         calculated.                                                              

Example 12 Cloning of HCMV gB in Poxvirus Vectors

Cloning of the HCMV gB gene into vaccinia donor plasmid, pMP22BHP. The 4800 bp HindIII-BamHI fragment of the HindIII D fragment of the HCMV DNA (Towne strain) was cloned into the 2800 bp HindIII-BamHI fragment of the plasmid pIBI24 (International Biotechnologies, Inc., New Haven, Conn.). By in vitro mutagenesis (Kunkel, 1985) using the oligonucleotides CMVM5 (SEQ ID NO:74) (5'-GCCTCATCGCTGCTGGATATCCGTTAAGTTTGTATCGTAATGGAATCCAGGATCTG-3') and CMVM3 (SEQ ID NO:75) (5"-GACAGAGACTTGTGATTTTTATAAGCTTCGTAAGCTGTCA-3'), the gB gene was modified to be expressed under the control of the vaccinia H6 promoter (Taylor et al., 1988a,b; Perkus et al., 1989). The plasmid containing the modified gB was designated 24CMVgB (5+3). The DNA sequence of the CMVgB gene is shown in FIG. 12 (SEQ ID NO:37).

Plasmid pMP2VCL (containing a polylinker region with vaccinia sequences upstream of the KIL host range gene) was digested within the polylinker with HindIII and XhoI and ligated to annealed oligonucleotides SPHPRHA A through D generating SP131 containing a HindIII site, H6 promoter -124 through -1 (Perkus et al., 1989) and a polylinker region. SPHPRHA A (SEQ ID NO:76) (5'-AGCTTCTTTATTCTATACTTAAAAAGTGAAAATAAATACAAAGGTTCTTGAGGGT-3') SPHPRHA B (SEQ ID NO:77) (5'-TGTGTTAAATTGAAAGCGAGAAATAATCATAAATTATTTCATTATCGCGATATCCGTTAA GTTTGTATCGTAC-3') SPHPRHA C (SEQ ID NO:78) (3'-TTATTAGTATTTAATAAAGTAATAGCGCTATAGGCAATTCAAACATAGCATGAGCT-5') SPHPRHA D (SEQ ID NO:79) (3'-AGAAATAAGATATGAATTTTTCACTTTTATTTATGTTTCCAAGAACTCCCAACACAATTT AACTTTCGCTCT-5').

The 2900 bp EcoRV-BamHI fragment of 24CMVgB (5+3) was cloned into the 3100 bp EcoRV-BglII fragment of SP131. This cloning step put the gB gene under the control of the H6 promoter. The resulting plasmid was designated SP131CMVgB.

Plasmid pSD22-H contains a 2.9 kb BglII fragment derived from the HindIII F region of the WR strain of vaccinia virus ligated into the BamHI site of pUC8. The unique BamHI site in pSD22-H is a nonessential site used as an insertion locus for foreign genes (Panicali and Paoletti, 1982). Plasmid pMP22BHP is a derivative of pSD22-H in which the unique BamHI site was modified by the addition of an expanded polylinker region for the insertion of foreign DNA. Plasmid pMP22BHP was digested with HindIII and ligated to a 2.9 kb HindIII fragment from SP131CMVgB (containing the H6 promoted gB gene) generating plasmid SAg22CMVgB. To modify the polylinker region in sAg22CMVgB, the plasmid was digested with BamHI followed by partial digestion with HindIII and purified. Ligation to a 50 bp BamHI/HindIII polylinker derived from IBI24 resulted in plasmid 22CMVgB.

Cloning of the HCMVgB gene into NYVAC donor plasmid pSD542. Plasmid pSD542 (a NYVAC TK locus donor plasmid) was derived from plasmid pSD513 (Tartaglia et al., 1992). The polylinker region in pSD513 was modified by cutting with PstI/BamHI and ligating to annealed synthetic oligonucleotides MPSYN288 (SEQ ID NO:80) (5'-GGTCGACGGATCCT-3') and MPSYN289 (SEQ ID NO:81) (5'-GATCAGGATCCGTCGACCTGCA-3') resulting in plasmid pSD542.

22CMVgB was digested with BamHI and NsiI to generate a fragment containing the H6 promoter and part of the gB gene, and with NsiI and PstI to generate a fragment containing the remainder of the gB gene. These two fragments were ligated to pSD542 that had been digested with BamHI and PstI within its' polylinker creating the NYVAC donor plasmid 542CMVgB. The DNA sequence of the CMVgB gene and flanking sequences contained in 542CMVgB is shown in FIGS. 13A and B (SEQ ID NO:38).

Cloning of the HCMV gB gene into the ALVAC donor plasmid CP3LVQH6. An 8.5 kb canarypox BglII fragment was cloned in the BamHI site of PBS-SK plasmid vector (Stratagene, La Jolla, Calif.) to form pWW5. Nucleotide sequence analysis revealed a reading frame designated C3 initiated at position 1458 and terminated at position 2897 in the sequence in FIGS. 14A-C (SEQ ID NO:39). In order to construct a donor plasmid for insertion of foreign genes into the C3 locus with the complete excision of the C3 open reading frame, PCR primers were used to amplify the 5' and 3' sequences relative to C3. Primers for the 5' sequence were RG277 (SEQ ID NO:82) (5'-CAGTTGGTACCACTGGTATTTTATTTCAG-3') and RG278 (SEQ ID NO:83) (5'-TATCTGAATTCCTGCAGCCCGGGTTTTTATAGCTAATTAGTCAAATGTGAGTTAATATTA G -3').

Primers for the 3" sequences were RG279 (SEQ ID NO:84) (5'-TCGCTGAATTCGATATCAAGCTTATCGATTTTTATGACTAGTTAATCAAATAAAAAGCAT ACAAGC-3') and RG280 (SEQ ID NO:85) (5'-TTATCGAGCTCTGTAACATCAGTATCTAAC-3'). The primers were designed to include a multiple cloning site flanked by vaccinia transcriptional and translational termination signals. Also included at the 5'-end and 3'-end of the left arm and right arm were appropriate restriction sites (Asp718 and EcoRI for left arm and EcoRI and SacI for right arm) which enabled the two arms to ligate into Asp718/SacI digested pBS-SK plasmid vector. The resultant plasmid was designated as pC3I.

A 908 bp fragment of canarypox DNA, immediately upstream of the C3 locus was obtained by digestion of plasmid pWW5 with NsiI and SspI. A 604 bp fragment of canarypox DNA was derived by PCR (Engelke et al., 1988) using plasmid PWW5 as template and oligonucleotides CP16 (SEQ ID NO:86) (5'-TCCGGTACCGCGGCCGCAGATATTTGTTAGCTTCTGC-3') and CP17 (SEQ ID NO:87) (5'-TCGCTCGAGTAGGATACCTACCTACTACCTACG-3'). The 604 bp fragment was digested with Asp718 and XhoI (sites present at the 5' ends of oligonucleotides CP16 and CP17, respectively) and cloned into Asp718-XhoI digested and alkaline phosphatase treated IBI25 (International Biotechnologies, Inc., New Haven, Conn.) generating plasmid SPC3LA. SPC3LA was digested within IBI25 with EcoRV and within canarypox DNA with NsiI and ligated to the 908 bp NsiI-SspI fragment generating SPCPLAX which contains 1444 bp of canarypox DNA upstream of the C3 locus.

A 2178 bp BglII-StyI fragment of canarypox DNA was isolated from plasmids pXX4 (which contains a 6.5 kb NsiI fragment of canarypox DNA cloned into the PstI site of pBS-SK). A 279 bp fragment of canarypox DNA was isolated by PCR (Engelke et al., 1988) using plasmid pXX4 as template and oligonucleotides CP19 (SEQ ID NO:88) (5'-TCGCTCGAGCTTTCTTGACAATAACATAG-3') and CP20 (SEQ ID NO:89) (5'-TAGGAGCTCTTTATACT ACTGGGTTACAAC-3'). The 279 bp fragment was digested with XhoI and SacI (sites present at the 5' ends of oligonucleotides CP19 and CP20, respectively) and cloned into SacI-XhoI digested and alkaline phosphatase treated IBI25 generating plasmid SPC3RA.

To add additional unique sites to the polylinker, pC3I was digested within the polylinker region with EcoRI and ClaI, treated with alkaline phosphatase and ligated to kinased and annealed oligonucleotides CP12 (SEQ ID NO:90) (5'-AATTCCTCGAGGGATCC-3') and CP13 (SEQ ID NO:91) (5'-CGGGATCCCTCGAGG-3') (containing an EcoRI sticky end, XhoI site, BamHI site and a sticky end compatible with ClaI) generating plasmid SPCP3S. SPCP3S was digested within the canarypox sequences downstream of the C3 locus with StyI and SacI (pBS-SK) and ligated to a 261 bp BglII-SacI fragment from SPC3RA and the 2178 bp BglII-StyI fragment from pXX4 generating plasmid CPRAL containing 2572 bp of canarypox DNA downstream of the C3 locus. SPCP3S was digested within the canarypox sequences upstream of the C3 locus with Asp718 (in PBS-SK) and AccI and ligated to a 1436 bp Asp718-AccI fragment from SPCPLAX generating plasmid CPLAL containing 1457 bp of canarypox DNA upstream of the C3 locus. CPLAL was digested within the canarypox sequences downstream of the C3 locus with StyI and SacI (in pBS-SK) and ligated to a 2438 bp StvI-SacI fragment from CPRAL generating plasmid CP3L containing 1457 bp of canarypox DNA upstream of the C3 locus, stop codons in six reading frames, early transcription termination signal, a polylinker region, early transcription termination signal, stop codons in six reading frames, and 2572 bp of canarypox DNA downstream of the C3 locus.

The early/late H6 vaccinia virus promoter (Taylor et al., 1988a,b; Perkus et al., 1989) was derived by PCR (Engelke et al., 1988) using pRW838 (a plasmid containing the rabies glycoprotein gene (Kieny et al., 1984) linked to the H6 promoter) as template and oligonucleotides CP21 (SEQ ID NO:92) (5'-TCGGGATCCGGGTTAATTAATTAGTTATTAGACAAGGTG-3') and CP22 (SEQ ID NO:93) (5'-TAGGAATTCCTCGAGTACGATACAAACTTAAGCGGATATCG-3'). The PCR product was digested with BamHI and EcoRI (sites present at the 5' ends of oligonucleotides CP21 and CP22, respectively) and ligated to CP3L that was digested with BamHI and EcoRI in the polylinker generating plasmid VQH6CP3L.

ALVAC donor plasmid VQH6CP3L was digested within the polylinker with XhoI and within the H6 promoter with NruI and ligated to a NruI/HindIII fragment from 22CMVgB containing part of the H6 promoter and gB gene and a polylinker derived from pIBI24 by XhoI and HindIII digestion generating the ALVAC donor plasmid CP3LCMVgB. The DNA sequence of the CMVgB gene plus additional flanking DNA sequences in plasmid CP3LCMVgB is shown in FIGS. 15A-C (SEQ ID NO:40).

Cloning of the HCMV gB gene deleted of its transmembrane region into the NYVAC donor plasmid pSD553. Plasmid pSD553 is a vaccinia deletion/insertion plasmid of the COPAK series. It contains the vaccinia K1L host range gene (Gillard et al., 1986; Perkus et al., 1990) within flanking Copenhagen vaccinia arms, replacing the ATI region (ORFs A25L, A26L; Goebel et al., 1990a,b). pSD553 was constructed as follows.

Left and right vaccinia flanking arms were constructed by polymerase chain reaction (PCR) using pSD414, a pUC8-based clone of vaccinia SalI B (Goebel et al., 1990a,b) as template. The left arm was synthesized using synthetic deoxyoligonucleotides MPSYN267 (SEQ ID NO:94) (5'-GGGCTGAAGCTTGCTGGCCGCTCATTAGACAAGCGAATGAGGGAC-3') and MPSYN268 (SEQ ID NO:95) (5'-AGATCTCCCGGGCTCGAGTAATTAATTAATTTTTATTACACCAGAAAAGACGGCTTGAGA T C-3') as primers. The right arm was synthesized using synthetic deoxyoligonucleotides MPSYN269 (SEQ ID NO:96) (5'-TAATTACTCGAGCCCGGGAGATCTAATTTAATTTAATTTATATAACTCATTTTTTGAATA T ACT-3') and MPSYN270 (SEQ ID NO:97) 5'-TATCTCGAATTCCCGCGGCTTTAAATGGACGGAACTCTTTTCCCCC-3') as primers. The two PCR-derived DNA fragments containing the left and right arms were combined in a further PCR reaction. The resulting product was cut with EcoRI/HindIII and a 0.9 kb fragment isolated. The 0.9 kb fragment was ligated with pUC8 cut with EcoRI/HindIII, resulting in plasmid pSD541. The polylinker region located at the vaccinia ATI deletion locus was expanded as follows. pSD541 was cut with BglII/XhoI and ligated with annealed complementary synthetic oligonucleotides MPSYN333 (SEQ ID NO:98) (5'-GATCTTTTGTTAACAAAAACTAATCAGCTATCGCGAATCGATTCCCGGGGGATCCGGTAC CC-3') and MPSYN334 (SEQ ID NO:99) (5'-TCGAGGGTACCGGATCCCCCGGGAATCGATTCGCGATAGCTGATTAGTTTTTGTTAACAA A A-3') generating plasmid pSD552. The K1L host range gene was isolated as a 1 kb BglII (partial)/HpaI fragment from plasmid pSD452 (Perkus et al., 1990). pSD552 was cut with BglII/HpaI and ligated with the K1L containing fragment, generating pSD553.

A HindIII fragment from SP131CMVgB (containing the HCMVgB gene under the control of the H6 promoter) was filled in with the klenow fragment of DNA polymerase I and ligated into plasmid pSD553 which had been SmaI digested and alkaline phosphatase treated. The resulting NYVAC donor plasmid (in which the H6 promoted gB is in the same orientation as K1L) was designated 553H6CMVgB. The DNA sequence of the CMVgB gene plus additional flanking DNA sequences in plasmid 553H6CMVgB is shown in FIGS. 16A and B (SEQ ID NO:41).

The sequence of CMVgB deleted of its transmembrane region is presented in FIG. 17 (SEQ ID NO:42). The nucleotides encoding the transmembrane region were deleted in the following manner. Oligonucleotides SPgB3 (SEQ ID NO:100) (5'-GATCCATGGACTCGACAGCGGCGTCTCTGCATGCAGCCGCTGCAGA-3') and SPgB4 (SEQ ID NO:101) (5'-AGCTTCTGCAGCGGCTGCATGCAGAGACGCCGCTGTCGAGTCCATG-3') were kinased, annealed and cloned into BamHI/HindIII digested and alkaline phosphatase treated IBI24 generating plasmid SPCMVgB2. Oligonucleotides SPgB1 (SEQ ID NO:102) (5'-TACGAATTCTGCAGTTCACCTATGACACGTTGC-3') and SPgB2 (SEQ ID NO:103) (5'-ATAGGATCCATGGTCGTCCAGACCCTTGAGGTAGGGC-3') were used in PCR with plasmid SP131CMVgB as template to generate a 0.7 kb fragment. This fragment was digested with EcoRI/BamHI and cloned into EcoRI/BamHI digested and alkaline phosphase treated IBI24 generating plasmid SPCMVgB1. A 0.7 kb EcoRI/NcoI fragment from SPCMVgB1 was ligated to EcoRI/NcoI digested and phosphatase treated SPCMVgB2 generating plasmid SPCMVgB3. The unique NcoI site in SPCMVgB3 was deleted by mutagenesis (Mandecki, 1986) using oligonucleotide SPgB5 (SEQ ID NO:104) 5'-GCCCTACCTCAAGGGTCTGGACGACACTCGACAGCGGCGTCTCTGCAT-3') generating plasmid SPCMVgB4. A 0.7 kb PstI fragment from SPCMVgB4 was ligated to a 6.6 kb PstI fragment from 553H6CMVgB generating NYVAC donor plasmid 553H6CMVgBTM⁻. This plasmid contains the gB gene under the control of the H6 promoter with its transmembrane region deleted (amino acids 715-772; Spaete et al., 1988). The DNA sequence of the transmembrane deleted CMVgB gene plus additional flanking DNA sequences in plasmid 553H6CMVgBTM⁻ is shown in FIGS. 18A and B (SEQ ID NO:43).

Clonina the HCMVgB gene deleted of its transmembrane region and containing an altered cleavage site into NYVAC donor plasmid pSD553. The sequence of CMVgB deleted of its transmembrane region and containing an altered cleavage site is presented in FIG. 19 (SEQ ID NO:44). The alteration of the cleavage site was accomplished in the following manner. Oligonucleotides SPgB8 (SEQ ID NO:105) (5'-AATTGGTGACCG-3') and SPgB9 (SEQ ID NO:106) (5'-GATCCGGTCACC-3') were kinased, annealed and cloned into EcoRI/BamHI digested and alkaline phosphatase treated IBI24 generating plasmid BstIBI. A 1.4 kb BstEII/SpHI fragment from 553H6CMVgBTM⁻ was cloned into BstEII/SpHI digested and alkaline phosphatase treated BstIBI generating plasmid SPCMVgB5.

Oligonucleotides SPgB10 (SEQ ID NO:107) (5'-TGAAAGACCGAATTCTGCGT-3') plus SPgB11 (SEQ ID NO:108) (5'-TGCGATTCATCGGTTTGTTGTAGAT-3') and SPgB12 (SEQ ID NO:109) (5'-GACCCTTGAGGTAGGGCGGC-3') plus SPgB13 (SEQ ID NO:110) (5'-ACTCATAATAGAACCATAAGATCTACAGATGGCAACAAT-3') were used in PCR with plasmid 553H6CMVgBTM⁻ to generate 0.7 and 0.8 kb fragments. These two fragments were combined in a PCR with oligonucleotides SPgB10 plus SPgB12 to generate a 1.2 kb fragment. The 1.2 kb fragment was digested with EcoRI and PstI and a 0.5 kb fragment isolated and cloned into EcoRI/PstI digested and alkaline phosphatase treated IBI24 generating plasmid SPCMVgB6. The 0.5 kb EcoRI/PstI fragment from SPCMVgB6 was used to replace the corresponding fragment in SPCMVgB5 generating plasmid SPCMVgB7. A 1.4 kb BstEII/SpHI fragment from SPCMVgB7 was used to replace the corresponding fragment in 553H6CMVgB generating NYVAC donor plasmid 553H6gBC⁻ TM⁻. This plasmid contains the gB gene under the control of the H6 promoter with its transmembrane region deleted (amino acids 715-772) and an alteration at the cleavage site (RTKR*ST modified to RTIRST where the asterisk indicated where cleavage normally occurs (Spaete et al., 1988) the S codon was modified to create a BglII restriction site). The DNA sequence of the cleavage site altered and transmembrane deleted CMVgB gene plus additional flanking DNA sequences in plasmid 553H6gBC⁻ TM⁻ is shown in FIGS. 20A and B (SEQ ID NO:45).

Example 13 Construction of Recombinant Poxviruses Containing HCMVgB

Procedures for transfection of recombinant donor plasmids into tissue culture cells infected with a rescuing poxvirus and identification of recombinants by Ln situ hybridization on nitrocellulose filters have been described (Guo et al., 1989; Panicali and Paoletti, 1982; Piccini et al., 1987; Perkus et al., 1993). Plasmid 542CMVgB was transfected into NYVAC (vP866) infected Vero cells (ATCC CCL#81) to generate the recombinant vP1001 (NYVAC-gB). Plasmid CP3LCMVgB was transfected into ALVAC infected primary chicken embryo fibroblast (CEF) cells to generate the recombinant vCP139 (ALVAC-gB). Plasmids 553H6CMVgB, 553H6CMVgBTM⁻ and 553H6gBC⁻ TM⁻ were transfected into NYVAC infected Vero cells to generate the recombinants vP1126, vP1128 and vP1145, respectively. Plasmid 22CMVgB was transfected into Vero cells infected with the WR L variant vaccinia virus (Panicali et al., 1981) to generate the recombinant vP992.

Example 14 Immunoprecipitation of HCMVgB Expressed by Poxvirus Recombinants

Immunoprecipitation assays were performed as described previously (Taylor et al., 1990) using gB specific guinea pig polyclonal serum (Gonczol et al., 1990). The apparent molecular weights of the gB specific bands corresponded to previously published results (Britt and Auger, 1986; Britt and Vugler, 1989; Reis et al., 1993). The intracellular fraction from vP992, vP1001, vCP139, vP1126, vP1128 and vP1145 contained a major band of apparent molecular weight 130-140 kDa, identifiable as the glycosylated uncleaved gB precursor. Fainter bands at approximately 110 kDa and 55 kDa, representing the N-terminal and C-terminal processed fragments were also seen in the cell fractions. The extracellular medium from vP1128 and vP1145 infected cells contained the uncleaved precursor and N-terminal and C-terminal processed fragments.

Example 15 Humoral Response of Laboratory Animals Inoculated with ALVAC-gB and NYVAC-gB

Following a single immunization of CBA mice with vP1001 (NYVAC-gB), neutralizing antibody titers of the sera of inoculated mice were assessed (Gonczol et al., 1986). Antibodies capable of neutralizing HCMV were detected (Table 21) in the sera of mice 14-21 days later (geometric mean titers of 1:16) and between 28-60 days post-immunization (gmt=1:26). A single immunization of CBA mice with vCP139 (ALVAC-gB) generated HCMV neutralizing antibody titers of 1:64 gmt (14-21 days pi) and 1:111 gmt (between 28 and 60 days pi). Thus, immunization of mice with NYVAC and ALVAC recombinants expressing HCMV gB elicited antibodies able to neutralize the infectivity of HCMV.

ALVAC-gB (vCP139) was evaluated for safety and immunogenicity in human volunteers. After two inoculations with 10⁶.3 TCID₅₀ of this recombinant, no serious reactions were noted.

                  TABLE 21                                                         ______________________________________                                         HCMV Neutralizing Antibodies in CBA mice                                                Days After Immunization                                               Immunization                                                                              14-21        21-28   28-60                                          ______________________________________                                         NYVAC-gB   16                                                                             16                                                                                                  32                                                                             24                                                                             32                                                                             24                                             ALVAC-gB   32                                                                             64                                                                             128                                                                            64                                                                                          64                                                                             128                                                                                    128                                                                            96                                             ______________________________________                                          Immunization was i.p. with 2-4 × 10.sup.8 PFU of recombinant             viruses.                                                                 

Guinea pigs were immunized twice with ALVAC-gB (days 0 and 28) and sera were tested for the presence of HCMV neutralizing antibody. HCMV neutralizing antibody was detected (Table 22) in the sera on day 34 (gmt=60), day 42 (gmt=60) and day 56(gmt=60). Thus, immunization of guinea pigs with ALVAC-gB elicited antibodies able to neutralize the infectivity of HCMV.

                  TABLE 22                                                         ______________________________________                                         HCMV Neutralizing Antibodies in Guinea Pigs                                    Inoculated with ALVAC-gB                                                                Days                                                                  Guinea Pig #                                                                              0     14       28  34     42  56                                    ______________________________________                                         19         <4    <4       <4  64     64  64                                    20         <4    <4       <4  32     64  64                                    21         <4    <4       <4  12     32  64                                    22         <4    <4       <4  48     48  32                                    23         <4    <4       4   96     46  46                                    24         <4    <4       <4  46     46  32                                    ______________________________________                                          Guinea pigs were inoculated by intramuscular route on days 0 and 28 with       10.sup.6.3 TCID.sub.50                                                   

Example 16 Cloning of HCMVgH in Poxvirus Vectors

Cloning of the HCMVgH gene into the NYVAC donor plasmid pSD550. The HCMVgH gene was isolated from genomic DNA (Towne strain) by PCR using oligonucleotides SPgH1 (SEQ ID NO:111) (5'-TATCTGCAGATGCGGCCAGGCCTCCCCTCCTAC-3') and SPgH2 (SEQ ID NO:112) (5'-CCGAAGCTTTCAGCATGTCTTGAGCATGC-3'). The resulting 2.3 kb fragment was digested with PstI (site at the 5' end of SPgH1) and HindIII (site at the 5' end of SPgH2) and cloned into PstI/HindIII digested and alkaline phosphatase treated IBI24 generating plasmid SPgH1. The sequence of CMVgH is presented in FIG. 21 (SEQ ID NO:46).

The 3' end of the gH gene in SPgH1 was modified to contain a vaccinia virus early transcription termination signal (Yuen and Moss, 1987) and a unique XhoI restriction site in the following manner. SPgH1 was digested within the 3' end of the gH gene with SpHI and within IBI24 with HindIII and the fragment containing gH was purified and ligated to kinased and annealed oligonucleotides SPgH16 (SEQ ID NO:113) (5'-CTCAAGACATGCTGATTTTTATCTCGAGA-3') and SPgH17 (SEQ ID NO:114) (5'-AGCTTCTCGAGATAAAAATCAGCATGTCTTGAGCATG-3') generating plasmid SPgH2.

Kinased and annealed oligonucleotides SPgH12 (SEQ ID NO:115) (5'-AATTCTCGAGTTTATTGGGAAGAATATGATAATATTTTGGGATTTC-3'), SPgH13 (SEQ ID NO:116) (5'-AAAATTGAAAATATATAATTACAATATAAAATGCGGCCCGGG-3'), SPgH14 (SEQ ID NO:117) (5'-GATCCCCGGGCCGCATTTTATATTGTAATTATAT-3') and SPgH15 (SEQ ID NO:118) (5'-ATTTTCAATTTTGAAATCCCAAAATATTATCATATTCTTCCCAATAAACTCGAG-3') were ligated to EcoRI/BamHI digested and alkaline phosphatase treated IBI24 generating plasmid SPgH3 which contains a unique XhoI site, the entomopox 42K promoter and nucleotide sequences encoding the first four amino acids of HCMVgH (underlined bases in codons three and four in oligonucleotides SPgH13 (SEQ ID NO:116) and SPgH14 (SEQ ID NO:117) were modified to create a SmaI site without altering the amino acid sequence). Oligonucleotides SPgH18 (SEQ ID NO:119) (5'-TTAGAATTCCCCGGGCTCCCCTCCTACCTCATCGT-3') and SPgH19 (SEQ ID NO:120) (5'-TTACTGCAGTAAGTGTTAAGTCTCTGTTGGTATC-3') were used in PCR with plasmid SPgH1 as template to derive a 0.4 kb fragment. This fragment was digested with SmaI and PstI and cloned into SmaI/PstI digested and alkaline phosphatase treated SPgH3 generating plasmid SPgH5 which contains a unique XhoI site, the 42K promoter and 5' 15% of the HCMVgH gene. A 0.4 kb EcoRI/BglII fragment from SPgH5 was ligated to a 4.7 kb EcoRI/BglII fragment from SPgH3 generating plasmid SPgH6 which contains the 42K promoted gH gene flanked by XhoI sites.

Plasmid pSD550 (an I4L locus donor plasmid) was derived from plasmid pSD548 (Tartaglia et al., 1992). The polylinker region in pSD548 was modified by cutting with BglII and SmaI and ligating to annealed synthetic oligonucleotides 539A (SEQ ID NO:121) (5'-AGAAAAATCAGTTAGCTAAGATCTCCCGGGCTCGAGGGTACCGGATCCTGATTAGTTAAT T TTTGT-3') and 539B (SEQ ID NO:122) (5'-GATCACAAAAATTAACTAATCAGGATCCGGTACCCTCGAGCCCGGGAGATCTTAGCTAAC T GATTTTTCT-3') resulting in plasmid pSD550. The 2.3 kb XhoI fragment from SPgH6 was cloned into XhoI digested and alkaline phosphatase treated pSD550 generating the NYVAC donor plasmid I4L42KgH in which the orientation of gH is in the same direction as the replaced I4L gene. The DNA sequence of CMVgH plus additional flanking DNA sequences in plasmid I4L42KgH are shown in FIGS. 22A and B (SEQ ID NO:47).

Cloning of the HCMVgH gene into the ALVAC donor plasmid NVQC5LSP. A C5 insertion vector containing 1535 bp upstream of C5, polylinker containing KpnI/SmaI/XbaI and NotI sites and 404 bp of canarypox DNA (31 base pairs of C5 coding sequence and 373 bp of downstream sequence) was derived in the following manner. A genomic library of canarypox DNA was constructed in the cosmid vector puK102 (Knauf and Nester, 1982) probed with pRW764.5 (a PuC9 based plasmid containing an 880 bp canarypox PvuII fragment which includes the C5 ORF Nucleotides 1372 to 2251 in FIG. 8 (SEQ ID NO:27)) and a clone containing a 29 kb insert identified (pHCOS1). A 3.3 kb ClaI fragment from pHCOS1 containing the C5 region was identified. The C5 open reading frame is initiated at position 1537 and terminated at position 1857 in the sequence shown in FIG. 8 (SEQ ID NO:27).

The C5 insertion vector was constructed in two steps. The 1535 bp upstream sequence was generated by PCR amplification using oligonucleotides C5A (SEQ ID NO:123) (5'-ATCATCGAATTCTGAATGTTAAATGTTATACTTTG-3') and C5B (SEQ ID NO:124) (5'-GGGGGTACCTTTGAGAGTACCACTTCAG-3') and purified genomic canarypox DNA as template. This fragment was digested with EcoRI (within oligoC5A) and cloned into EcoRI/SmaI digested pUC8 generating C5LAB. The 404 bp arm was generated by PCR amplification using oligonucleotides C5C (SEQ ID NO:125) (5'-GGGTCTAGAGCGGCCGCTTATAAAGATCTAAAATGCATAATTTC-3') and C5DA (SEQ ID NO:126) (5'-ATCATCCTGCAGGTATTCTAAACTAGGAATAGATG-3'). This fragment was digested with PstI (within oligoC5DA) and cloned into SmaI/PstI digested C5LAB generating pC5L.

pC5L was digested within the polylinker with Asp718 and NotI, treated with alkaline phosphatase and ligated to kinased and annealed oligonucleotides CP26 (SEQ ID NO:127) (5'-GTACGTGACTAATTAGCTATAAAAAGGATCCGGTACCCTCGAGTCTAGAATCGATCCCGG GTTTTTATGA CTAGTTAATCAC-3') and CP27 (SEQ ID NO:128) (5'-GGCCGTGATTAACTAGTCATAAAAACCCGGGATCGATTCTAGACTCGAGGGTACCGGATC C TTTTTATAGCTAATTAGTCAC-3') (containing a disabled Asp718 site, translation stop codons in six reading frames, vaccinia early transcription termination signal (Yuen and Moss, 1987), BamHI KpnI XhoI XbaI ClaI and SmaI restriction sites, vaccinia early transcription termination signal, translation stop codons in six reading frames, and a disabled NotI site) generating plasmid C5LSP. The polylinker region in C5LSP was further modified by digesting with BamHI and ligating to annealed oligonucleotides CP32 (SEQ ID NO:129) (5'-GATCTTAATTAATTAGTCATCAGGCAGGGCGAGAACGAGACTATCTGCTCGTTAATTAAT T AGGTCGACG-3') and CP33 (SEQ ID NO:130) (5'-GATCCGTCGACCTAATTAATTAACGAGCAGATAGTCTCGTTCTCGCCCTGCCTGATGACT A ATTAATTAA-3') generating plasmid VQC5LSP. VQC5LSP was digested with EcoRI, treated with alkaline phosphatase, ligated with kinased and annealed oligonucleotide CP29 (SEQ ID NO:131) (5'-AATTGCGGCCGC-3') and digested with NotI. The linearized plasmid was purified and self ligated to generate plasmid NVQC5LSP. The 2.3 kb XhoI fragment from SPgH6 was cloned into XhoI digested and alkaline phosphatase treated NVQC5LSP generating the ALVAC donor plasmid NVQC5L42KgH in which the orientation of gH is in the same direction as the deleted C5 gene. The DNA sequence of CMVgH plus additional flanking DNA sequences in plasmid NVQC5L42KgH are shown in FIGS. 23A and B (SEQ ID NO:27).

Cloning of the HCMVQH gene into the vaccinia donor plasmid pSD157K1LINS. Plasmid. pHK (which contains the WR vaccinia HindIII K fragment cloned in pBR322) was digested with HindIII/BglII and a 1.2 kb fragment isolated and cloned into BamHI/HindIII digested pBS-SK⁺ yielding plasmid pBS-HKARM. pBS-HKARM was digested with Asp718 in the polylinker region, blunt ended with the klenow fragment of E. Coli DNA polymerase, and digested with HindIII at the pBS/vaccinia junction. The resulting 4.1 kb vector fragment was ligated to a 2.0 kb NruI/HindIII fragment from pHM-1 (pHM-1 contains the WR vaccinia virus HindIII M fragment cloned in pBR322) resulting in plasmid PMPWRMK. pMPWRMK was cut with HpaI and ligated with annealed synthetic oligonucleotides MPSYN527 (SEQ ID NO:132) (5'-ATAAAAATTAGCTACTCAGGTACCCTGCAGTCGCGAGGATCCGAATTCCCCGGGCTCGAG T GATTAATTAGTTTTTAT-3') and MPSYN528 (SEQ ID NO:133) (5'-ATAAAAACTAATTAATCACTCGAGCCCGGGGAATTCGGATCCTCGCGACTGCAGGGTACC T GAGTAGCTAATTTTTAT-3'). The resulting plasmid is pSD157K1LINS. pSD157K1LINS was digested within its polylinker region with XhoI, treated with alkaline phosphatase and ligated to the 2.3 kb XhoI fragment from SPgH6 yielding plasmid MP804-42KgH (which contains the HCMVgH gene and vaccinia K1L gene both in the same orientation.) The DNA sequence of CMVgH plus additional flanking DNA sequences in plasmid MP804-42KgH are shown in FIG. 24 (SEQ ID NO:49).

Example 17 Construction of Recombinant Poxviruses Containing HCMVgH

Plasmid I4L42kgH was transfected into NYVAC infected CEF cells to generate the recombinant vP1173 (containing HCMVgH). The same plasmid was transfected into vP1001 infected Vero cells to generate the recombinant vP1183 (containing HCMVgB and gH).

Plasmid NVQC5L42KgH was transfected into ALVAC infected CEF cells to generate the recombinant vCP236 (containing HCMVgH). The same plasmid was transfected into vCP139 infected CEF cells to generate the recombinant vCP233 (containing HCMVgB and gH). Vaccinia virus vP1170 (which contains Ecogpt under the transcriptional control of the entomopoxvirus 42K promoter in place of the deleted K1L gene) was used to infect Vero cells transfected with plasmid MP804-42KgH to generate the recombinant vP1205B.

Example 18 Immunoprecipitation of HCMVgH Expressed by Poxvirus Recombinants

Immunoprecipitation performed with a monoclonal antibody specific for HCMVgH demonstrated the expression of an 86 kDa gH protein (Pachl et al., 1989) by recombinants vP1173, vP1183, vP1205B, vCP233 and vCP236. Immunoprecipitation with the gB specific guinea pig polyclonal serum demonstrated correct expression of gB by recombinants vP1183 and vCP233.

The HCMV 72-kDa immediate early 1 protein (IE1) is a target for CD8⁺ cytotoxic T cells in humans (Borysiewicz et al., 1988) and is recognized by CD4⁺ T cells (Alp et al., 1991). For one individual the peptide specificities of proliferative and MHC-class I-restricted cytotoxic determinants on IE1 were determined and found to be spatially distinct segments of the exon 4 coding region (Alp et al., 1991).

The IE1 protein has been shown to up-regulate expression from its own promoter (Cherrington and Mocarski, 1989) as well as expression from the HIV LTR (Biegalke and Geballe, 1991; Ghazal et al., 1991) and expression of the promoters for the cellular genes c-myc, c-fos and hsp70 (Hagemeier et al., 1992; Santomenna and Colberg-Poley, 1990; Colberg-Poley et al., 1992). Lafemina et al., (1989) reported that the IE1 protein expressed in stable cell lines preferentially associates with metaphase chromosomes and proposed that this protein may be involved in maintenance of a putative plasmid state for HCMV DNA during latency.

In the following Examples 19-30, the development of poxvirus recombinants expressing the entire IE1 gene, IE1 deleted of amino acids 2-32, IE1 deleted of amino acids 292-319 or the exon 4 segment of IE1 are provided. These studies were performed in order to develop a form of the IE1 gene product that would be incapable of translocation to the nucleus, thus decreasing its potential to act as a transactivator, while maintaining its ability to be recognized by CD8⁺ cytotoxic T cells. Example 45 demonstrates that an ALVAC recombinant expressing an altered form of the IE1 protein (deleted of amino acids 2-32) which unlike the full length gene product is found in both the nucleus and cytomplasm of infected cells, can re-stimulate cytotoxic effector cells from HCMV seropositive individuals.

Example 19 Cloning of the Entire HCMV IE1 Gene in Poxvirus Vectors

Cloning of the HCMV IE1 gene into the vaccinia donor plasmid pSD22-H. The entire HCMV IE1 gene (AD169 strain) was derived as a 1.5 kb fragment by PCR using plasmid pJD083 as template (Akrigg et al., 1985) along with oligonucleotides IE3 (SEQ ID NO:134) (5'-ACGGATCCATAAAAATTACTGGTCAGCCTTGCTTC-3') and IE5 (SEQ ID NO:135) (5'-ATCCGTTAAGTTTGTATCGTAATGGAGTCCTCTGCCAAGAGA-3'). The DNA sequence of CMV IE1 is presented in FIG. 25 (SEQ ID NO:50). Plasmid pSD486H6340 (which contains an irrelevant gene linked precisely to H6 promoter) was digested (within the H6 promoter) with NruI and (at the 3' end of the irrelevant gene) with BamHI and ligated to the BamHI digested 1.5 kb PCR fragment (BamHI site located at the 5' end of oligonucleotide IE3) generating plasmid pSD486H6HCMVIE1.

The H6 promoted IE1 gene was obtained from pSD486H6HCMVIE1 as a 1.6 kb fragment by digestion with BamHI followed by partial BglII digestion and ligated to BamHI digested pSD22-H yielding plasmid pSD22-HCMVIE1. The DNA sequence of CMV IE1 plus additional flanking DNA sequences in plasmid pSD22-HCMVIE1 are shown in FIG. 26 (SEQ ID NO:51).

Cloning of the HCMVIE1 gene into the vaccinia donor plasmid pSD554. Oligonucleotides SPIE1 (SEQ ID NO:136) (5'-CGCGAATTCTCGCGATATCCGTTAAGTTTGTATCGTAATGGAGT-3') and SPIE2 (SEQ ID NO:137) (5'-GCCTCTAGAGTTAACCTCCTTCCTCAACAT-3') were used in PCR with plasmid pSD486H6HCMVIE1 as template to generate a 181 bp fragment. This fragment was digested with EcoRI and XbaI and cloned into EcoRI/XbaI digested and alkaline phosphatase treated IBI24 generating plasmid SPIE1 containing part of the H6 promoter and the first 135 bp of the IE1 gene. Oligonucleotides SPIE3 (SEQ ID NO:138) (5'-CGGTCTAGAGGTTATCAGTGTAATGAAGC-3') and SPIE4 (SEQ ID NO:139) (5'-CCGAAGCTTCTCGAGATAAAAATTACTGGTCAGCCTTGCTTCTAGT-3') were used in PCR with plasmid pSD486H6HCMVIE1 as template to generate a 506 bp fragment. This fragment was digested with XbaI and HindIII and cloned into XbaI/HindIII digested and alkaline phosphatase treated IBI24 generating plasmid SPIE2 containing the 3' end of the IE1 gene, a vaccinia early transcription termination signal and an XhoI site. SPIE1 was digested at the 3' end of the inserted fragment of the IE1 gene with HindII and within the IBI24 polylinker with HindIII, alkaline phosphatase treated and ligated to a 903 bp HindII-BglII fragment from pSD486H6HCMVIE1 and a 464 bp BqlII-HindIII fragment from SPIE2 generating plasmid SPIE3 containing the entire IE1 gene linked to part of the H6 promoter.

Plasmid pSD553 was cut with NruI and ligated with a SmaI/NruI fragment containing the synthetic H6 promoter (Perkus et al., 1989) upstream from the NruI site located at -26 relative to the translation initiation codon. The resulting plasmid, pMP553H6, was digested with NruI and BamHI and ligated to annealed oligonucleotides MPSYN347 (SEQ ID NO:140) (5'-CGATATCCGTTAAGTTTGTATCGTAATCTGCAGCCCGGGGGGG-3') and MPSYN348 (SEQ ID NO:141) (5'-GATCCCCCGGGCTGCAGATTACGATACAAACTTAACGGATATCG-3'). The resulting plasmid, pSD554, contains the entire H6 promoter region through nucleotide -1 relative to the initiation codon, followed by a polylinker region. pSD554 was digested with NruI and XhoI and ligated to a 1.5 kb NruI/XhoI fragment from SPIE3 generating plasmid COPAKH6IE. The DNA sequence of CMV IE1 plus flanking DNA sequences in plasmid COPAKH6IE are shown in FIGS. 27A and B (SEQ ID NO:52).

Example 20 Construction of Recombinant Poxviruses Containing the Entire HCMVE1 Gene

Plasmid pSD22-HCMVIE1 was transfected into Vero cells infected with the WR L variant to generate the recombinant vP893. Plasmid COPAKH6IE was transfected into NYVAC infected Vero cells to generate the recombinant vP1161.

Example 21 Expression of the Entire IE1 Gene by Poxvirus Recombinants

Immunoprecipitation studies performed with a monoclonal antibody specific for HCMVIE1 demonstrated the expression of a 72 kDa IE1 protein (Blanton and Tevethia, 1981; Cameron and Preston, 1981) by recombinants vP893 and vP1161. Immunofluorescence studies (performed as described in Taylor et al., 1990) revealed nuclear localization of the IE1 gene product.

Example 22 Cloning of the HCMVEI1 Gene (Lacking Amino Acids 292-319) into the Vaccinia Donor Plasmid pSD554

The DNA sequence of CMVIE1 lacking amino acids 292-319 is shown in FIG. 28 (SEQ ID NO:53). This deletion was made in the following manner. Plasmid SPIE3 was digested with SpeI and a 4239 bp fragment isolated (which lacks nucleotides 868-958 encoding amino acids 292-319). This fragment was self ligated generating plasmid SPIE4. A 1.4 kb NruI/XhoI fragment from SPIE4 was ligated to NruI/XhoI digested pSD554 generating plasmid COPAKH6IEN⁻. The DNA sequence of CMVIE1 lacking amino acids 292-319 plus flanking DNA sequences in plasmid COPAKH6IEN⁻ are shown in FIGS. 29A and B (SEQ ID NO:54).

Example 23 Construction of a Recombinant Poxvirus Containing the HCMV Ie1 Gene Lacking Amino Acids 292-319

Plasmid COPAKH6IEN⁻ was transfected into NYVAC infected Vero cells to generate the recombinant vP1160.

Example 24 Expression of the HCMVIE1 Gene Lacking Amino Acids 292-319

Immunoprecipitation assays demonstrated the expression of a 69 kDa protein in cells infected with vP1160 consistent with the deletion of amino acids 292-319. Immunofluorescence studies revealed nuclear localization of this gene product.

Example 25 Cloning of the Exon 4 Segment of HCMVIE1 in Poxvirus Vectors

Cloning of the Exon 4 segment of HCMVIE1 in NYVAC donor plasmid SPI4LH6. The DNA sequence of the Exon 4 segment of HCMVIE1 is shown in FIG. 30 (SEQ ID NO:55). This segment of the gene was obtained in the following manner. Oligonucleotides SPIE5 (SEQ ID NO:142) (5'-CGCGAATTCTCGCGATATCCGTTAAGTTTGTATCGTAATGAAACAGATTAAGGTTCGAGT -3') and SPIE6 (SEQ ID NO:143) (5'-GCCTCTAGATGCCGCCATGGCCTGACT-3') were used in PCR with plasmid pSD486H6HCMVIE1 to generate a 0.5 kb fragment. This fragment was digested with EcoRI and XbaI and cloned into EcoRI/XbaI digested and alkaline phosphatase treated IBI24 generating plasmid SPIE5. Plasmid SPIE3 was digested with EcoRI and NcoI and a 3.6 kb fragment purified and ligated to a 0.47 kb EcoRI-NcoI fragment from SPIE5 generating plasmid SPIE6 which contains the Exon 4 segment of IE1 linked to part of the H6 promoter.

The early/late H6 vaccinia virus promoter (Guo et al., 1989; Perkus et al., 1989) was derived by PCR using PRW823 as template (a plasmid containing the H6 promoter linked to an irrelevant gene) and oligonucleotides CP30 (SEQ ID NO:144) (5'-TCGGGATCCGGGTTAATTAATTAGTCATCAGGCAGGGCG-3') and CP31 (SEQ ID NO:145) (5'-TAGCTCGAGGGTACCTACGATACAAACTTAACGGATATCG-3'). The PCR product was digested with BamHI and XhoI (sites present at the 5' end of CP30 and CP31, respectively) and ligated to BamHI/XhoI digested C5LSP generating plasmid VQH6C5LSP. This plasmid was used as template in PCR with oligonucleotides CP31 and RUB1 (SEQ ID NO:146) (5'-TCGGGATCCTTCTTTATTCTATACTTA-3'). The PCR product was digested with BamHI and XhoI (site present at the 5' ends of RUB1 and CP31, respectively) and ligated to BamHI/XhoI digested pSD550 generating plasmid SPI4LH6. A 1.3 kb NruI/XhoI fragment isolated from SPIE6 was cloned into NruI/XhoI digested and alkaline phosphatase treated SPI4LH6 generating plasmid I4LH6IE-Ex4 (in which the H6 promoted IE1 Exon 4 gene is in the same orientation as the replaced I4L gene). The DNA sequence of the Exon 4 segment of HCMVIE1 plus flanking DNA sequences in plasmid I4LH6IE-Ex4 are shown in FIG. 31 (SEQ ID NO:56).

Cloning of the Exon 4 fragment of HCMVIE1 in ALVAC donor plasmid NVQH6C5LSP. Plasmid VQH6C5LSP was digested with EcoRI, treated with alkaline phosphatase, ligated with kinased and annealed oligonucleotide CP29 and digested with NotI. The linearized plasmid was purified and self ligated generating plasmid NVQH6C5LSP. The 1.3 kb NruI/XhoI fragment from SPIE6 was cloned into NruI/XhoI digested and alkaline phosphatase treated NVQH6C5LSP generating plasmid NVQH6IE-Ex4 (in which the H6 promoted IE1 Exon 4 gene is in the same orientation as the replaced C5 gene). The DNA sequence of the Exon 4 segment of HCMVIE1 plus flanking DNA sequences in plasmid NVQH6IE-Ex4 are shown in FIG. 32A and B (SEQ ID NO:57).

Example 26 Construction of Recombinant Poxviruses Containing the Exon 4 Segment Of IE1

Plasmid I4LH6IE-Ex4 was transfected into NYVAC infected CEF cells to generate the recombinant vP1186. Plasmid NVQH6IE-Ex4 was transfected into ALVAC infected CEF cells to generate the recombinant vCP244.

Example 27 Expression of the Exon 4 Segment of HCMVIE1 by Poxvirus Recombinants

Immunofluorescence experiments revealed cytoplasmic localization of the IE-Exon 4 protein expressed by recombinants vP1186 and vCP244. Immunoprecipitation experiments with a monoclonal antibody specific for IE-Exon 4 demonstrated the expression of a 60 kDa protein in cells infected with vCP244 consistent with the predicted size of the exon 4 segment. Immunoprecipitation with a polyclonal rabbit serum raised against a bacterial Exon 4 fusion protein revealed the expression of a 60 kDa protein in cells infected with vP1186 and VCP244.

Example 28 Cloning of the HCMVIE1 Gene (Lacking Amino Acids 2-32) in Poxvirus Vectors

Cloning of the HCMVIE1 gene (lacking amino acids 2-32) in NYVAC donor plasmid SPI4LH6. The DNA sequence of HCMVIE1 lacking amino acids 2-32 is shown in FIG. 33 (SEQ ID NO:58). This segment was obtained in the following manner. Oligonucleotides SPIE9 (SEQ ID NO:147) 5'-AATTCTCGCGATATCCGTTAAGTTTGTATCGTAATGACGACGTTCCTGCAGACTATGTTG A GGAAGGAGGTT-3') and SPIE10 (SEQ ID NO:148) (5'-AACCTCCTTCCTCAACATAGTCTGCAGGAACGTCGTCATTACGATACAAACTTAACGGAT ATCGC GAG-3') were kinased, annealed and ligated to a 4.2 kb HindII/EcoRI digested and alkaline phosphatase treated fragment from SPIE3 generating plasmid SPIE8. A 1.4 kb NruI/XhoI fragment from SPIE8 (containing part of the H6 promoter and IE1 lacking amino acids 2-32) was ligated to NruI/XhoI digested and alkaline phosphatase treated SPI4LH6 generating plasmid I4LH6IEd32. The DNA sequence of HCMVIE1 lacking amino acids 2-32 plus flanking DNA sequences in plasmid I4LH6IEd32 are shown in FIG. 34 (SEQ ID NO:59).

Cloning of the HCMVIE1 gene (lacking amino acids 2-32) in ALVAC donor plasmid NVQH6C5LSP. The 1.4 kb NruI/XhoI fragment from SPIE8 was cloned into NruI/XhoI digested and alkaline phosphatase treated NVQH6C5LSP generating plasmid NVQH6IEd32. The DNA sequence of HCMVIE1 lacking amino acids 2-32 plus flanking DNA sequences in plasmid NVQH6IEd32 are shown in FIGS. 35A and B (SEQ ID NO:60).

Example 29 Construction of Poxvirus Recombinants Containing the IE1 Gene Lacking Amino Acids 2-32

Plasmid I4LH6IEd32 was transfected into NYVAC infected CEF cells to generate the recombinant vP1201. Plasmid NVQH6IEd32 was transfected into ALVAC infected CEF cells to generate the recombinant vCP256.

Example 30 Expression of IE1 Lacking Amino Acids 2-32 by Poxvirus Recombinants

Immunofluorescence experiments revealed both nuclear and cytoplasmic localization of the IE1 protein lacking amino acids 2-32 by recombinants vP1201 and vCP256. Immunoprecipitation with a polyclonal rabbit serum raised against a bacterial exon 4 fusion protein revealed the expression of a 68 kDa protein in cells infected with vP1201 consistent with the predicted size.

Example 31 Cloning of the HCMV pp65 Gene in Poxvirus Vectors

Cloning of the HCMV pp65 gene in NYVAC donor plasmid SPHA-H6. pSD456 is a subclone of Copenhagen vaccinia DNA containing the HA gene (A56R; Goebel et al., 1990a,b) and surrounding regions. pSD456 was used as a template in PCR for synthesis of left and right vaccinia arms flanking the A56R ORF. The left arm was synthesized using oligonucleotides MPSYN279 (SEQ ID NO:149) (5'-CCCCCCGAATTCGTCGACGATTGTTCATGATGGCAAGAT-3') and MPSYN280 (SEQ ID NO:150) (5'-CCCGGGGGATCCCTCGAGGGTACCAAGCTTAATTAATTAAATATTAGTATAAAAAGTGAT TTATTTTT-3'). The right arm was synthesized using oligonucleotides MPSYN281 (SEQ ID NO:151) (5'-AAGCTTGGTACCCTCGAGGGATCCCCCGGGTAGCTAGCTAATTTTTCTTTTACGTATTAT A TATGTAATAAACGTTC-3') and MSYN312 (SEQ ID NO:152) (5'-TTTTTTCTGCAGGTAAGTATTTTTAAAACTTCTAACACC-3'). The purified PCR fragments for the left and right arms were combined in a further PCR reaction. The resulting product was digested with EcoRI/HindIII. The resulting 0.9 kb fragment was cloned into EcoRI/HindIII digested pUC8 resulting in plasmid pSD544.

pSD544 was digested within its polylinker with XhoI, filled in with klenow and treated with alkaline phosphatase. Plasmid SP126 (equivalent to SP131) was digested with HindIII, treated with klenow and the H6 promoter isolated by digestion with SmaI. Ligation of the H6 promoter fragment to pSD544 generated SPHA-H6.

The HCMV pp65 gene was PCR amplified using HCMV genomic DNA as template (Towne strain) and oligonucleotides pp651 (SEQ ID NO:153) (5'-GATTATCGCGATATCCGTTAAGTTTGTATCGTAATGGCATCCGTACTGGGTCCCATTTCG GG-3') and pp651R (SEQ ID NO:154) (5'-GCATAGGTACCGGATCCATAAAAATCAACCTCGGTGCTTTTTGGGCG-3').The DNA sequence of CMVpp65 is shown in FIG. 36 (SEQ ID NO:61). The 1.6 kb product was digested with NruI and BamHI (site present at the 5' end of oligonucleotides pp651 and pp651R, respectively) and cloned into NruI/BamHI digested SPHA-H6 generating plasmid CMV65.1. This plasmid contained the pp65 gene linked to the H6 promoter, however, the first 30 bp of the pp65 gene were missing.

To derive a plasmid containing the first 30 bp of the pp65 gene oligonucleotides RNApp65I (SEQ ID NO:155) (5'-TAGTTCGGATCCCCGCTCAGTCGCCTACA-3') and pp65R4 (SEQ ID NO:156) (5'-ATCAAGGGATCCATCGAAAAAGAAGAGCG-3') were used in PCR with genomic DNA. The resulting 1 kb fragment was digested with BamHI (BamHI sites present at the 5' ends of both oligonucleotides) and cloned into BamHI digested IBI24 generating plasmid pp65.7. Plasmid pp65.7 was used in PCR with oligonucleotides pp651B (SEQ ID NO:157) (5'-GATTATCGCGATATCCGTTAAGTTTGTATCGTAATGGAGTCGCGCGGTCGCCGTTGTCCC G -3') and pp65BstXI (SEQ ID NO:158) (5'-ACCTGCATCTTGGTTGC-3') to generate a 0.5 kb fragment. This fragment was digested with NruI and BstXI (sites at the 5' ends of oligonucleotides pp651B and pp65BstXI, respectively) and ligated to a 4.8 kb NruI/BstXI fragment of CMV65.1 generating plasmid pCMV65.2. This plasmid contains the entire pp65 gene linked precisely to the H6 promoter oriented in the same direction as the replaced HA gene. The DNA sequence of CMVpp65 plus flanking DNA sequences in plasmid pCMV65.2 are shown in FIG. 37 (SEQ ID NO:62).

Cloning of the HCMV pp65 gene in ALVAC donor plasmid pMPC616E6VQ. FIGS. 38A and B (SEQ ID NO:63) is the sequence of a 3.7 kb segment of canarypox DNA. Analysis of the sequence revealed a reading frame designate C6L initiated at position 377 and terminated at position 2254. A C6 insertion vector containing 370 bp upstream of C6, polylinker containing SmaI, PstI, XhoI and EcoRI sites, and 1156 bp of downstream sequence was derived in the following manner. The 0.4 bp upstream sequence was generated by PCR amplification of a cosmid clone derived from purified genomic canarypox DNA using oligonucleotides C6A1SG (SEQ ID NO:159) (5'-ATCATCGAGCTCGCGGCCGCCTATCAAAAGTCTTAATGAGTT-3') and C6B1SG (SEQ ID NO:160) (5'-GAATTCCTCGAGCTGCAGCCCGGGTTTTTATAGCTAATTAGTCATTTTTTCGTAAGTAAG T ATTTTATTTAA-3'). The 1.2 kb downstream arm was generated by PCR amplification of the same template using oligonucleotides C6C1SG (SEQ ID NO:161) (5'-CCCGGGCTGCAGCTCGAGGAATTCTTTTTATTGATTAACTAGTCAAATGAGTATATATAA T TGAAAAAGTAA-3') and C6D1SG (SEQ ID NO:162) (5'-GATGATGGTACCTTCATAAATACAAGTTTGATTAAACTTAAGTTG-3'). These fragments were fused by a third PCR employing gel purified 0.4 and 1.2 kb fragments as template for primers C6A1SG (SEQ ID NO:159) and C6D1SG (SEQ ID NO:162). The resulting 1.6 kb fragment was isolated from an agarose gel, digested with SacI and KpnI and ligated to similarly digested pBS generating C6 insertion plasmid pC6L.

Plasmid pMPC616E6VQ was derived by cloning a HpaI-XhoI fragment containing the H6 promoter precisely linked to an irrelevant gene into Sma-XhoI digested pC6L. pMPC616E6VQ was digested with NruI and BamHI and the 4 kb vector fragment (NruI-BamHI) and 0.6 kb C6 flanking arm fragment (BamHI-BamHI) isolated. These two fragments were combined in a ligation with a 1.7 kb NruI-BamHI fragment from pCMV65.2 (containing part of the H6 promoter linked to the p65 gene) generating plasmid CMV65C6.1 which contained a C6 flanking arm, H6 promoter and the pp65 gene but lacked the 0.6 kb C6 flanking arm. CMV65C6.1 was digested with BamHI, treated with alkaline phosphatase and ligated to the 0.6 kb C6 flanking arm generating plasmid CMV65C6.2 in which C6 flanking arms are present on both sides of the H6-pp65 insert. The DNA sequence of CMVpp65 plus flanking DNA sequences in plasmid CMV65C6.2 are shown in FIGS. 39A and B (SEQ ID NO:64).

Cloning of the HCMVpp65 gene into the vaccinia donor plasmid pSD157 K1LINS. Plasmid pCMV65.2 was digested with KpnI, treated with Mung Bean Nuclease and digested with BamHI generating a 1.7 kb fragment containing H6-pp65. PSD157K1LINS was digested with BamHI and SmaI and ligated to the 1.7 kb fragment generating plasmid CMV65.WR. The DNA sequence of CMVpp65 plus flanking DNA sequences in plasmid CMV65.WR are shown in FIG. 40 (SEQ ID NO:65).

Example 32 Construction of Recombinant Poxviruses Containing HCMVpp65

Plasmid pCMV65.2 was transfected into NYVAC infected Vero cells to generate the recombinant vP1184 (containing HCMVpp65), into vP1001 infected Vero cells to generate the recombinant vP1196 (containing HCMVgB and pp65) and into vP1183 infected Vero cells to generate the recombinant vP1210 (containing HCMVgB, gH and pp65).

Plasmid CMV65C6.2 was transfected into ALVAC infected CEF cells to generate the recombinant vCP260 (containing HCMVpp65).

Plasmid CMV65.WR was transfected into vP1170 infected Vero cells to generate the recombinant vP1214 (WR-pp65).

Example 33 Expression of HCMVpp65 by Poxvirus Recombinants

Immunoprecipitation experiments with a monoclonal antibody specific for HCMV pp65 demonstrated the expression of a 65 kDa protein (Pande et al., 1991) by recombinants vP1184, vP1214, vCP260, vP1196 and vP1210. In addition, immunoprecipitation with gB specific guinea pig polyclonal sera demonstrated correct expression of gB by recombinants vP1196 and vP1210 and immunoprecipitation with a gH specific monoclonal antibody demonstrated correct expression of gH by recombinant vP1210.

Example 34 Cloning of the HCMV pp150 Gene in Poxvirus Vectors

Cloning of the pp150 gene into the NYVAC donor plasmid pSD541. The DNA sequence of CMVpp150 is shown in FIG. 41 (SEQ ID NO:66). Oligonucleotides pp150.1B (SEQ ID NO:163) (5'-TTCGGATCCGGTTCTGGAGAAAAGCC-3') and pp150R6 (SEQ ID NO:164) (5'-GCTTCCAAGCTTTCCTGAAGGGATTGTAAGCC-3') were used in PCR with Towne genomic DNA to generate a 2 kb fragment from the 5' end of pp150. This fragment was digested with BamHI and HindIII and cloned into BamHI/HindIII digested and alkaline phosphatase treated IBI24 generating plasmid pp150.5.

Oligonucleotides pp150.9 (SEQ ID NO:165) (5'-TTCGGATCCGGCTTTCAGTCTCGTCTCC-3') and pp150END2 (SEQ ID NO:166) (5'-TTCGGATCCATGCAATTGCCCGCGGACAAC-3') were used in PCR with Towne DNA to generated a 1.8 kb fragment which includes the 3' end of the gene. This fragment was digested with BamHI and cloned into BamHI digested and alkaline phosphatase treated PUC8 yielding pp150.3.

Oligonucleotides SP150-3 (SEQ ID NO:167) (5'-TTCGAATTCGCTAGCTTTATTGGGAAGAATATGATAATATTTTGGGATTTCAAAATTGAA A ATATATAATTACAATATAAAATGAGTTTGCAGTTTATC-3') and SP150-4 (SEQ ID NO:168) (5'-TTCTCTAGATGAGCTCGTTGAACAGCAC-3') were used in PCR with plasmid pp150.5 as template to generate a 259 bp fragment. This fragment was digested with EcoRI and XbaI and cloned into EcoRI/XbaI digested and alkaline phosphatase treated IBI24 generating plasmid 150.5MP. This plasmid contains a NheI site, 65 bp entomopoxvirus 42K promoter and bases 1-170 from the 5' end of the pp150 gene. The underlined base in the sequence of oligonucleotide SP150-3 (position -53 of the promoter) is missing in this clone.

Oligonucleotides SP150-1 (SEQ ID NO:169) (5'-CCGAAGCTTGCTAGCAATAAAAACTATTCCTCCGTGTTCTTAAT-3') and SP150-2 (SEQ ID NO:170) (5'-GCCTCTAGATACGTAAAGCTAAGTTATC-3') were used in PCR with plasmid pp150.3 as template to generate a 907 bp fragment. This fragment was digested with XbaI and HindIII and cloned into XbaI/HindIII digested and alkaline phosphatase treated IBI24 yielding plasmid 150.3MP. This plasmid contains nucleotides 2273-3141 from pp150 followed by a vaccinia early transcription termination signal (T₅ ATT) (Yuen and Moss, 1987) and a NheI site. pp150 nucleotide 2748 (FIG. 41; SEQ ID NO:66) in this clone is an A not a C as in pp150.3, this change is silent.

Plasmid pp150.3 was digested with SnaBI and HindIII and a 3451 bp fragment isolated. Plasmid 150.3MP was digested with SnaBI and HindIII and 873 bp fragment isolated. Ligation of these two fragments yielded plasmid 150.3MC which contains pp150 nucleotides 1473-3141 followed by T₅ ATT and a NheI site.

Plasmid 150.5MP was digested with SacI and HindIII and a 3056 bp fragment isolated. Plasmid pp150.5 was digested with SacI and HindIII and a 1816 bp fragment isolated. Ligation of these two fragments yielded plasmid 150.5MC which contains a NheI site, 65bp 42K promoter and pp150 nucleotides 1-1981.

Plasmid 150.5MC was digested with HpaI and HindIII and a 4634 bp fragment isolated. Plasmid 150.3MC was digested with HpaI and HindIII and a 1412 bp fragment isolated. Ligation of these two fragments yielded plasmid 150.1 which contains a NheI site, 65bp 42K promoter, nucleotides 1-3141 pp150, T₅ ATT and a NheI site.

Plasmid pSD541 is a vaccinia insertion plasmid which is deleted for vaccinia sequences encompassing the A25L and A26L ORFs (Goebel et al., 1990a,b). The deletion junction consists of a polylinker region containing XhoI, SmaI and BglII restriction sites, flanked on both sides by stop codons and early vaccinia transcriptional terminators (Yuen and Moss, 1987). pSD541 was constructed by polymerase chain reaction (PCR) using cloned vaccinia SalI E plasmid pSD414 as template. Synthetic oligonucleotides MPSYN267 (SEQ ID NO:94) (5'-GGGCTCAAGCTTGCGGCCGCTCATTAGACAAGCGAATGAGGGAC-3') and MPSYN268 (SEQ ID NO:95) (5'-AGATCTCCCGGGCTCGAGTAATTAATTAATTTTTATTACACCAGAAAAGACGGCTTGAGA T C-3') were used as primers to generate the left vaccinia arm and synthetic oligonucleotides MPSYN269 (SEQ ID NO:96) (5'-TAATTACTCGAGCCCGGGAGATCTAATTTAATTTAATTTATATAACTCATTTTTTGAATA T ACT-3') and MPSYN270 (SEQ ID NO:97) (5'-TATCTCGAATTCCCGCGGCTTTAAATGGACGGAACTCTTTTCCCC-3') were used to generate the right vaccinia arm. PCR products consisting of the left and right vaccinia arms were combined, and subjected to PCR amplification. The PCR product was digested with EcoRI and HindIII and electrophoresed on a agarose gel. The 0.8 kb fragment was isolated and ligated into pUC8 cut with EcoRI/HindIII, resulting in plasmid pSD541.

Plasmid pSD541 was digested in its polylinker region with SmaI and alkaline phosphatase treated. Plasmid 150.1 was digested with NheI, treated with klenow and a 3224bp fragment (containing 42K-pp150) isolated. Ligation of these two fragments yielded plasmid 150.7. The DNA sequence of CMVpp150 plus flanking DNA sequences in plasmid 150.7 are shown in FIGS. 42A and B (SEQ ID NO:67).

Cloning of the pp150 gene into ALVAC donor plasmid PMM117. Plasmid PMM117 is a derivative of pC6L with a modified polylinker region. PMM117 was digested in its polylinker with EcoRI filled in with klenow and treated with alkaline phosphatase. Plasmid 150.1 was digested with NheI, treated with klenow and a 3224bp fragment (containing 42K-pp150) isolated. Ligation of these two fragments generated plasmid 150.6. The DNA sequence of CMVpp150 plus flanking DNA sequences in plasmid 150.6 are shown in FIGS. 43A and B (SEQ ID NO:68).

Cloning of the pp150 gene into vaccinia donor plasmid pSD157K1LINS. Plasmid pSD1571LINS was digested in its polylinker region with SmaI and alkaline phosphatase treated. Plasmid 150.1 was digested with NheI, treated with klenow and a 3224 bp fragment (containing 42K-pp150) isolated. Ligation of these two fragments generated plasmid 150.4. The DNA sequence of CMVpp150 plus flanking DNA sequences in plasmid 150.4 are shown in FIGS. 44A and B (SEQ ID NO:69).

Example 35 Construction of Recombinant Poxviruses Containing HCMVpp150

Plasmid 150.4 was transfected into vP1170 infected CEF cells to generate the recombinant vP1238 (WR-pp150).

Plasmid 150.7 was transfected into NYVAC infected CEF cells to generate the recombinant vP1247 (NYVAC-pp150).

Plasmid 150.6 was transfected into ALVAC infected CEF cells to generate the recombinant vCP284 (ALVAC-pp150).

Example 36 Expression of HCMVpp150 by Poxvirus Recombinants

Western blot (Harlow and Lane, 1988) with a monoclonal antibody specific for HCMVpp150 demonstrated the expression of a 150 kDa protein in cells infected with vP1238 which comigrated with a protein present in HCMV infected cells. Expression of a 150 kDa protein was observed in vP1247 and vCP284 infected cells by immunoprecipitation with the pp150 specific monoclonal antibody.

Example 37 Developing a NYVAC Donor Plasmid Containing the HCMVgH and IE1 Exon 4 Genes

Plasmid I4LH6IE-Ex4 was linearized with BamHI, filled in with klenow and treated with alkaline phosphatase yielding a 4.9 kb fragment. Plasmid gH6-3 was digested with XhoI, filled in with klenow and a 2.3 kb fragment (containing 42K-gH) isolated. These two fragments were ligated to generate plasmid I4L42KgHH6IE-Ex4. The DNA sequence of CMVgH and IE-Exon4 plus additional flanking sequences in plasmid I4L42KgHH6IE-Ex4 are shown in FIGS. 45A and B (SEQ ID NO:70).

Example 38 Construction of NYVAC Recombinants Containing HCMVgB.⁺ gH.⁺ pp65.⁺ IE-EXON 4, HCMVgB.⁺ gh.⁺ pp65.⁺ pp150 or HCMVgB.⁺ gh.⁺ pp65.⁺ IE-EXON 4 and pp150

Plasmid I4L42KgHH6IE-Ex 4 was transfected into vP1196 infected Vero cells to generate the recombinant vP1216 (containing HCMVgB, gH, pp65, IE-Exon 4). Plasmid 150.7 was transfected into vP1216 infected CEF cells to generate the recombinant vP1251 (containing HCMVgB, gH, IE-Exon 4, pp65, pp150). Plasmid 150.7 was transfected into vP1210 infected Vero cells to generate the recombinant vP1262 (containing HCMV-gB, gH, pp65, pp150).

Example 39 Expression of the HCMV Genes in vP1216, vP1251, vP1262

Immunoprecipitation with monoclonal antibodies specific for gB, gH, pp65 and IE-Exon 4 demonstrated the correct expression of all four genes by recombinant vP1216. Immunoprecipitation with monoclonal antibodies specific for gB, gH, pp65 and IE-Exon 4 demonstrated the correct expression of these four genes by recombinant vP1251. Immunoprecipitation with monoclonal antibodies specific for gB, gH and pp65 demonstrated the correct expression of these three genes by recombinant vP1262. Western blot with a monoclonal antibody specific for pp150 demonstrated the correct expression of this gene by recombinants vP1251 and vP1262.

Example 40 Developing an ALVAC Donor Plasmid Containing the HCMV pp65 and pp150 genes

Plasmid CMV65C6.2 was linearized with EcoRI, filled in with klenow and treated with alkaline phosphatase generating a 6.3 kb fragment. Plasmid 150.1 was digested with NheI, filled in with klenow and a 3.2 kb fragment (42K-pp150) isolated. Ligation of these two fragments yielded plasmid 150.8. The DNA sequence of CMVpp65 and pp150 plus additional flanking sequences in plasmid 150.8 are shown in FIGS. 46A to C (SEQ ID NO:71).

Example 41 Construction of an ALVAC Recombinant Containing HCMVgB. gH, pp65 AND pp150

Plasmid 150.8 was transfected into vPC233 infected CEF cells to generate an ALVAC-gB, gH, pp65, pp150 recombinant (vCP280).

Example 42 Expression of the HCMV Genes in vCP280

Immunoprecipitation with monoclonal antibodies specific for gB, gH and pp65 demonstrated the correct expression of these three genes by recombinant vCP280.

Example 43 Cloning of HCMVgL in Poxvirus Vectors Deriving a NYVAC Donor Plasmid Containing gB and gL

Oligonucleotides UL115A (SEQ ID NO:171) (5'-GCCTCTAGAATGTGCCGCCGCCCGGATTGC-3') and UL115B (SEQ ID NO:172) (5'-CGCAAGCTTAGCGAGCATCCACTGCTTGAGGGC-3') were used in PCR with Towne DNA as template to generate a 853bp fragment. This fragment was digested with XbaI and HindIII and cloned into XbaI/HindIII digested and alkaline phosphatase treated IBI24 generating plasmid UL115.1. The sequence of CMVgL is presented in FIG. 47 (SEQ ID NO:72).

Oligonucleotides UL115M (SEQ ID NO:173) (5'-TCCAAGCTTAGATCTATAAAAATTAGCGAGCATCCACTGCTTGAGGGCCATAGC-3') and UL115N (SEQ ID NO:174) (5'-GCCTCTAGATGCTGACGCTGTTGAGCTCGGAC-3') were used in PCR with plasmid UL115.1 as template to generate a 498bp fragment. This fragment was digested with HindIII and XbaI and cloned into HindIII/XbaI digested and alkaline phosphatase treated IBI24 generating plasmid UL115.2.

Oligonucleotides UL115G2 (SEQ ID NO:175) (5'-CGCGAATTCTCGCGATATCCGTTAAGTTTGTATCGTAATGTGCCGCCGCCCGGATTGC -3') and UL115H2 (SEQ ID NO:176) (5'-GCCTCTAGATTCCAGCGCGGCGCTGTGTCCGAGC-3') were used in PCR with plasmid UL115.1 as template to generate a 450bp fragment. This fragment was digested with EcoRI and XbaI and cloned into EcoRI/XbaI digested and alkaline phosphatase treated IBI24 generating plasmid UL115.3.

Plasmid UL115.3 was digested with HindIII and SacI and a 3226bp fragment isolated. Plasmid UL115.2 was digested with HindIII and SacI and a 469bp fragment isolated. Ligation of these two fragments yielded plasmid UL115.4.

Plasmid UL115.4 was digested with NruI and BglII and a 865bp fragment isolated. Plasmid I4LH6 was digested with NruI and BglII and a 3683bp fragment isolated. Ligation of these two fragments yielded plasmid I4LH6gL.

To correct a one base deletion in the H6 promoter in I4LH6gL this plasmid was digested with EcoRV treated with alkaline phosphatase and a 3805bp fragment isolated. Plasmid I4LH6 was digested with EcoRV and a 736bp fragment isolated. Ligation of these two fragments yielded plasmid I4LH6CgL.

Plasmid 542CMVgB was linearized with BamHI and treated with alkaline phosphatase. Plasmid I4LH6CgL was digested with BamHI and BglII and a 968bp fragment (containing the H6 promoted gL gene) isolated. Ligation of these two fragments generated plasmid 542CMVgBgL. The DNA sequence of CMVgL and CMVgB plus additional flanking DNA sequences in plasmid 542CMVgBgL are shown in FIGS. 48A and B (SEQ ID NO:73).

Example 44 Developing a NYVAC Recombinant Containing gB, gH, gL, pp65, pp150, IE1-Exon 4 or gB, gH, gL, pp65, pp150

Plasmid 542CMVgBgL was transfected into vP1251 infected CEF cells to generate a NYVAC gB, gH, gL, pp65, pp150, IE1-Exon 4 recombinants (NYVAC-CMV6: vP1302 and vP1302B).

Plasmid 542CMVgBgL is transfected into vP1262 infected cells to generate NYVAC recombinant vP1312 (NYVAC-CMV5).

Example 45 Human Cytotoxic T Lymphocyte Responses to HCMV Proteins

Lymphocytes comprising the antigen-specific segment of the immune system may functionally react to antigen by producing antibodies (B-lymphocytes) or by becoming cytotoxic T lymphocytes (CD8+ T-lymphocytes). ALVAC recombinants expressing HCMV proteins that are known to be recognized by human cytotoxic T lymphocytes (CTLs) are capable of re-stimulating human cellular immune responses with characteristics of classical CTLs.

Thirteen individuals for which there was previously established EBV-transformed B-cell lines (LBCL) for use as CTL targets were screened for CTL responses to HCMV gB, IE1, and pp65. Although only one of these volunteer blood donors had an established clinical history of HCMV infection, seven were found to be HCMV seropositive by virtue of their sera containing antibodies which neutralized HCMV.

Stimulation of HCMV 1E1 CTLs by ALVAC-1E1 (vCP256): Whole blood was collected into heparinized Vacutainer tubes from each volunteer donor by venipuncture. The mononuclear cell fraction was separated from the remainder of the blood components by centrifugation over Leucoprep gradients, washed several times by centrifugation in Stim Medium (MEM containing 5% fetal bovine serum [FBS], 2 mM L-glutamine, 10⁻⁴ M 2-mercaptoethanol, 100 IU/ml penicillin, and 100 μg/ml streptomycin), counted for viable cells with trypan blue, and resuspended at 5×10⁶ cells/ml in Stim Medium (responder cells). A portion of the mononuclear cells were resuspended at 10⁷ cells/ml in MEM containing 2% FBS and infected with recombinant ALVAC expressing HCMV 1E1 (vCP256) at a multiplicity of infection of 25 for approximately 1 hour at 37 C. Following incubation, sufficient Stim Medium was added to dilute the infected cells to 5×10⁵ cells/ml (stimulator cells). Equal volumes of responder cells and stimulator cells were added to upright 25 cm² tissue culture flasks or to the wells of 24-well tissue culture plates and incubated in 5% CO₂ /95% air at 37 C. for 6 days. Target cells were prepared by infecting LBCLs with recombinant WR vaccinia virus expressing HCMV IE1 (vP893) similarly to the infection of stimulator cells except the target cells were incubated overnight at 4×10⁵ cells/ml in RPMI 1640 medium containing 20% FBS. Following incubation, the mononuclear cells and the target cells were washed by centrifugation in Assay Medium (RPMI 1640 medium containing 10% FBS, 2 mM Lglutamine, 5×10⁻⁵ M 2-mercaptoethanol, 100 IU/ml penicillin, and 100 μg/ml streptomycin). Target cells were incubated in Na₂ ⁵¹ CrO₄ for 1 hour, washed by centrifugation in Assay Medium, resuspended to 10⁵ cells/ml in Assay Medium, and held on ice until use. Following centrifugation, the mononuclear cells were diluted to 2×10⁶ cells/ml in Assay Medium. One tenth ml of mononuclear cells and 0.1 ml of ⁵¹ Cr labelled, infected target cells were added to the wells of 96-well round bottom tissue culture plates. These volumes and cell densities resulted in an effector to target ratio (E:T) of 20:1. The tissue culture plates were centrifuged at 250 g for 2 minutes and incubated in 5% CO₂ /95% air at 37 C. for 4 to 5 hours. Following incubation, 0.1 ml of supernatant fluid from each well was collected using Skatron filter wicks and counted for released radioactivity. Percent cytoxicity was calculated as: ##EQU1##

Maximum release was determined by the addition of 5% sodium dodecyl sulfate to target cells while spontaneous release was determined by incubating target cells in the absence of effector cells. In none of the experiments presented did spontaneous release of ⁵¹ Cr from target cells exceed 20% of maximum ⁵¹ Cr release.

Following in vitro stimulation with ALVAC recombinants expressing a single HCMV protein, mononuclear cells from four of the seven seropositive volunteer donors lysed autologous targets expressing HCMV IE1 (FIG. 49) and mononuclear cells from six of the seven seropositive donors lysed autologous targets expressing HCMV pp65 (FIG. 50). Re-stimulated mononuclear cells from none of the HCMV seropositive donors lysed autologous targets expressing HCMV gB.

The mononuclear cells from HCMV seronegative volunteer donors, when re-stimulated similarly to the mononuclear cells of the HCMV seropositive donors, failed to lyse autologous target cells expressing HCMV IE1 or HCMV pp65 (FIG. 49 and FIG. 50, respectively).

In all cases except one, the cytotoxic effector cells only lysed autologous, but not nonautologous, target cells expressing the appropriate HCMV protein. The single exception, mononuclear cells from Donor 7C, following re-stimulation with ALVAC pp65 (vCP260), was capable of lysing nonautologous target cells expressing HCMV pp65. However, it was later demonstrated that Donor 7C and the donor for the nonautologous target cell line share HLA-B7 of the human major histocompatibility complex (MHC).

Stimulation of HCMV IE1 CTLs by ALVAC-IE1 (vCP256): Human CTLs were stimulated in vitro and assayed for HCMV IE1 CTLs using similar methodology as in FIG. 49 except that following 6 days incubation for restimulation, the responder mononuclear cells were incubated with immunomagnetic beads coupled to monoclonal anti-human CD3, CD4, or CD8. Following incubation, the beads were removed by a magnet and therefore the CD3+, CD4+ or CD8+ cells. The cells adhering to the magnetic beads were uncoupled, washed and used in the cytotoxicity assay.

Representative of the phenotype of the cytotoxic responses of this HCMV seropositive cohort, the ALVAC-IE1 (vCP256) re-stimulated mononuclear cells from Donor 2A failed to lyse IE1-expressing targets following depletion of lymphocytes expressing CD3 and CD8, but not CD4 (FIG. 51). Furthermore, re-stimulated mononuclear cells that had been enriched for CD8, but not CD4, retained cytotoxic activity.

Thus, the cytotoxic effector cells derived from HCMV seropositive volunteer donors by re-stimulation in vitro with ALVAC recombinants expressing HCMV IE1 (vCP256) or HCMV pp65 (vCP260) were antigen specific, MHC-restricted, and expressed CD3 and CD8. These characteristics are consistent with those of classical cytotoxic T lymphocytes (CTLs).

These results show that ALVAC recombinants expressing HCMV proteins can serve as vaccines for the purpose of eliciting human cytotoxic T lymphocytes capable of mediating the destruction of HCMV-infected human cells. Furthermore, these data also show that these recombinant viruses can serve as reagents for the ex vivo stimulation and expansion of cytotoxic T lymphocyte clones for the purpose of immunotherapeutic applications (Riddell et al., 1992).

As discussed earlier, HCMV-gB can serve to elicit protective immunity in humans since 1) HCMV neutralizing antibody titer is reduced significantly when gB specific antibody is absorbed from human sera (Gonczol et al., 1991; Marshall et al., 1992) and 2) there is evidence for the activation of helper T cells by the gB protein in seropositive individuals (Liu et al., 1991). Gonczol et al., (1990) reported the immunoaffinity purified gB was immunogenic in human volunteers. In this study a single injection of the purified gB was able to induce high titers of HCMV neutralizing antibodies and lymphocyte proliferation in naturally seropositive individuals. In seronegative individuals three injections of the gB preparation induced transient HCMV neutralizing antibodies, a fourth injection induced a rapid reappearance and increase in titer of HCMV neutralizing antibodies.

These studies show the use of purified gB as a subunit vaccine. Additionally purified gB can also be used in prime/boost protocols in combination with NYVAC or ALVAC-gB recombinants. Recent studies have indicated that a prime/boost protocol, whereby immunization with a poxvirus recombinant expressing a foreign gene product is followed by a boost with a purified form of that gene product, elicits an enhanced immune response relative to the response elicited with either product alone. For example, humans immunized with a vaccinia recombinant expressing the HIV-1 envelope glycoprotein and boosted with purified HIV-1 envelope glycoprotein from a baculovirus recombinant exhibit higher HIV-1 neutralizing antibody titers than individuals immunized with just the vaccinia recombinant or purified envelope glycoprotein alone (Graham et al., 1993; Cooney et al., 1993). Humans immunized with two injections of ALVAC-HIV (vCP125) failed to develop HIV specific antibodies. Boosting with purified rgp160 from a vaccinia virus recombinant resulted in detectable HIV-1 neutralizing antibodies. Furthermore, specific lymphocyte T cell proliferation to rgp160 was clearly increased by the boost with rgp160. Envelope specific cytotoxic lymphocyte activity was also detected with this vaccination regimen (Pialoux et al., 1995). Macaques immunized with a vaccinia recombinant expressing the simian immunodeficiency virus (SIV) envelope glycoprotein and boosted with SIV envelope glycoprotein from a baculovirus recombinant are protected against a SIV challenge (Hu et al., 1991; 1992).

Example 46 Purification of HCMV Glycoprotein B

This Example involves purification of CMV glycoprotein B produced by a vaccinia recombinant, and the testing of its immunogenicity in laboratory animals in combination with ALVAC-CMV gB (vCP139).

COPAK recombinants vP1126, vP1128, and vP1145, each expressing a different form of gB, elicit CMV neutralizing antibodies in mice (Table 23) and therefore express gB in an immunogenic form. To select a virus and cell system, and an immunological reagent for CMV gB purification, gB expression by the three COPAK recombinants was compared by an immunoprecipitation assay, utilizing 5 different gB-specific monoclonal antibodies. Based on the assay results, a scheme was developed to purify gB from the medium of vP1145-infected VERO cells.

Immunoaffinity column bed material was prepared by crosslinking CMV gB-specific monoclonal antibody (mAb) CH380 to Protein A-agarose. This material was used to purify gB in a one-step procedure. Batches of gB were produced and evaluated for purity, as described in section III.

Immunoprecipitation Assay. Vero and HeLa cell monolayers in 60 mm dishes were infected with vP1126, vP1128, vP1145, or vP993 (described below) at an moi of 5 pfu/cell in serum-free medium. Medium and cells were harvested separately at 24 hours post infection. Immunoprecipitation (IP) assays were performed (Taylor et al., 1990) using the reagents described below, with rat anti-mouse IgG as a bridge to protein A for the monoclonals.

    ______________________________________                                         Virus:                                                                         vP1126:         COPAK-CMV gB (entire).                                                         Full length wild type gB                                       vP1128:         COPAK-CMV gB (TM.sup.-).                                                       Lacks transmembrane region                                     vP1145:         COPAK-CMV gB (TM.sup.-, Cl.sup.-                                               lacks transmembrane region                                                     and has an altered cleavage site.                              vP993:          COPAK control                                                  Reagents:                                                                      Guinea pig anti-CMV gB:                                                                        Obtained from Eva Gonczol (Wistar                                              Institute)                                                     Monoclonal CH380:                                                                              Obtained from PMs&v (Pereria and                                               Hoffman, 1986)                                                 Monoclonal 13-127                                                                              Advanced Biotechnologies, Inc.                                 Monoclonal 13-128                                                                              Advanced Biotechnologies, Inc.,                                                neutralizing, conformationally                                                 dependent                                                      Monoclonal HCMV-34                                                                             Cogent Diagnostics, neutralizing                               Monoclonal HCMV-37                                                                             Cogent Diagnostics, neutralizing                               Rabbit anti-p25 (Vaccinia E3L)                                                                 (obtained from Bert Jacobs,                                                    U. Arizona)                                                    ______________________________________                                    

Preparation of immunoaffinity chromatography bed material. One ml of immunoaffinity column bed material consisting of approximately 2.4 mg of mAb CH380 coupled to Protein A-agarose with the crosslinking agent dimethylpimelimidate was provided by Stephen Cockle, Connaught Laboratories, Limited (Willowdale, Ontario, Canada). mAb CH380 (Pereria and Hoffman, 1986) was used previously to purify CMV gB from a CMV viral envelope preparation (Gonczol et.al., 1990). The material from S. Cockle was used in preliminary experiments to further determine its utility in gB purification. To scale up gB production, additional bed material was prepared by the same method used by S. Cockle, as described below.

Preparation of monoclonal ch380. Four vials of lyophilized monoclonal CH380 (lot S1705, obtained from PMsv) were reconstituted in PBS (137 mM NaCl, 2.7 mM KCl, 1.5 mM KH₂ PO₄, 8.1 mM Na₂ HPO₄, pH 7.4) (1 ml each) and dialysed overnight versus PBS (final volume 3.5 ml). Protein concentration was determined to be 4.9 mg/ml by bicinchoninic acid assay (BCA assay, reagents obtained from Pierce, Rockford, Ill.). This preparation was then diluted in an equal volume of MAPS binding buffer (Bio-Rad cat# 153-6161; 31.4% w/v in milli-Q water, adjusted to pH 9, and filtered through a 22 mm membrane). To remove particulate material, the antibody preparation in MAPS buffer was centrifuged at 16,000×g for 30 min, and the protein concentration of the supernate was calculated from the absorbence at 280 nm, using 1.44 as the absorbence coefficient for IgG.

Preparation of protein a-agarose beads. Three ml of protein A-agarose beads (Bio-Rad cat # 153-6153) were washed 4 times with 2 volumes of MAPS binding buffer by gentle mixing in a closed tube and centrifugation for 5 min at 1000×g (1400 rpm in Beckman GPKR centrifuge, GH 3.7 rotor). The supernate was discarded after the last wash.

Bindina of monoclonal antibody to the beads. All of the mAb antibody from step 1 was added to the washed beads from step 2 and the mixture was rotated in a closed tube at 4° C. The amount of mAb bound to the beads was determined at 6-12 hour intervals by pelleting the beads (1000 g/5 min) and determining concentration of IgG in the supernatant by reading OD at 280 nm, as described above. Approximately 48 hour of incubation at 4° C. were required to reach 90% depletion of IgG from the supernate.

Covalent crosslinking of monoclonal antibody to the beads. After binding was 90% complete, the beads were washed 4 times with 6 ml (2 volumes) of 50 mM borate, 3M NaCl, pH9. The beads were then resuspended in 30 ml (10 volumes) of 200 mM borate, 3M NaCl, pH9, and the pH adjusted to 9±0.1. A sample of beads (100 μl) was removed for later evaluation of cross-linking. Cross linking reagent dimethylpimelimidate (DMP) was prepared immediately before use at a concentration of 500 mM in 200 mM borate, 3M NaCl, pH9. DMP was added to the beads to produce a final concentration of 20 mM, and the beads were mixed in a closed tube, end-over-end, for 30 min at room temperature. Another sample of beads (100 μl) was removed for evaluation of crosslinking. To quench residual crosslinking reagent, the beads were washed 2 times with 6 ml (2 volumes) of 200 mM ethanolamine, pH8 and then incubated in 30 ml (10 volumes) of 200 mM ethanolamine, pH8 by mixing end-over-end for 2 hours at room temperature. Finally the beads were washed 4 times with 6 ml (2 volumes) of PBS and stored in 6 ml of PBS with 0.01% NaN₃.

To determine the extent of crosslinking, the gel bead samples taken before and after DMP incubation were pelleted, supernates discarded, and the beads mixed with 2×SDS-PAGE sample buffer containing reducing agent. These samples were boiled and electrophoretically separated on a 10% polyacrylamide gel. After staining with Coomassie Blue, IgG heavy and light chains could be detected in the "before" samples, but not in the "after" samples, indicating good efficiency of crosslinking.

Based on protein concentration before and after incubation of the antibody with the beads, the resulting bed material was estimated to contain approximately 5 mg of monoclonal antibody per ml of protein A-agarose beads.

Purification of CMV gB by immunoaffinity column chromatography. Column buffers. PBS (137 mM NaCl, 2.7 mM KCl, 1.5 mM KH₂ PO₄, 8.1 mM Na₂ HPO₄), pH 7 (batch 1), pH 7.4 (batches 2-5), or pH 6.8 (batches 2-5); 0.1 M glycine, pH 2.5; 1 M tris, pH 8.5.

Columns. Column sizes varied from 0.3 to 4 ml volumes. When a new column was poured, it was stripped with 10 bed volumes (bv) of 0.1 M glycine, pH 2.5, followed by 10-20 bv of PBS, pH 7 or 7.4. At the end of each column run, the column was washed with at least 10 bv of PBS, pH 7. At the beginning of each run, it was washed again with at least 10 bv of PBS, pH 7. The columns were run at room temperature and, when not in use, stored at 4° C. in PBS+0.01% NaN₃.

Preparation of the crude gB sample. Roller bottles (850 cm²) were seeded with Vero cells in MEM+10% FBS. Medium was changed to serum-free MEM 2-12 hours before infection. Cells were infected with vP1145 at an MOI of 5 pfu/cell in a volume of 10 ml/RB of serum-free MEM. Virus was absorbed at 37° C. for 60 min and then 30 ml of serum-free MEM was added to each RB and incubation continued at 37° C. Medium was harvested at 16-24 hours post infection. The medium was clarified by centrifugation at 3000 rpm (Beckman GPKR centrifuge GH 3.7 rotor) for 15 min. The supernatant was recovered and further clarified by centrifugation at 20,000 rpm in a Beckman SW28 rotor for 60 min. The clarified medium was then concentrated (10 to 40-fold) by ultrafiltration with buffer exchange to PBS, pH 7.4, using one or more of the following ultrafiltration devices having 30,000 MWCO: Centricell-60 (Polysciences #19182-6), Centriprep-30 (Amicon #4306), or polysulfone immersible filter units (Polysciences #2250). This material was applied to the column as described below.

Column procedure. The crude gB sample was applied to the column at a flow rate of 0.03-0.09 ml/min, controlled by stopcock or peristaltic pump. After application of the sample, the column was washed at a flow rate of 0.2-0.6 ml/min with 10 bv PBS, pH7 (batch 1), or 20 bv of PBS, pH7.4 followed by 20 bv of PBS, pH6.8 (batches 2-5). Bound material was eluted with 10 bv of 0.1 M glycine, pH 2.5, collecting 500 μl (Batch 1,3) or 1 ml (batch 2,4,5) fractions into tubes containing 50 μl (Batch 1,3) or 100 μl (batch 2,4,5) of 1.0 M Tris, pH 8.5. One column (#28) was eluted with 0.1N glycine+0.1M Tris, pH7. CMV gB fractions were identified by SDS-PAGE on a 10% gel, under reducing conditions, followed by silver stain (Bio-Rad kit #161-0443).

Treatment of eluted gB. After identification by SDS-PAGE and silver stain the CMV gB fractions were pooled and concentrated in one of 2 ways: 1) Dialysis against 0.1×PBS and 10-fold vacuum concentration (majority of batch 1), or 2) Precipitation with 70% ammonium sulfate and resuspension in PBS. Protein concentration of the gB samples was determined by bicinchoninic acid microplate assay (BCA reagents from Pierce, Rockford, Ill.). Five batches of gB were prepared and frozen in aliquots at -70° C.

Evaluation of purified gB. Slot blot. Slot blot analysis was utilized to measure relative quantities of CMV gB in crude preparations, flow-through fractions, and elution fractions from affinity column purification. Serial two-fold dilutions in PBS were made of each test sample, and these were applied to nitrocellulose paper with the Schleicher and Scheull Manifold II slot blot apparatus. Each test included serially diluted samples of purified gB with a known protein concentration (determined by BCA microplate assay) as a standard. CMV gB was detected with monoclonal CH380 diluted 1:100 followed by 125I goat anti-mouse (NEN # NEX159, at 0.1 Ci/ml). Slot blot signals on the autoradiograph were scanned and analyzed by densitometry (PDI, Inc., Huntington Station, N.Y., Quantity One densitometer program). The amount of CMV gB in each test sample was determined by linear regression analysis as compared to a gB standard curve.

Western blot. Test samples were electrophoretically separated on a 10% gel under reducing conditions, and blotted onto nitrocellulose paper (Harlow and Lane, 1988). The blot was probed for the presence of CMVgB, mouse IgG, vaccinia, and Vero cell proteins with the following reagents:

    ______________________________________                                         ANTIGEN  PRIMARY ANTIBODY DETECTION                                            ______________________________________                                         CMV gB   Monoclonal CH380 .sup.125 I goat anti-mouse                                    diluted 1:100    (NEN # NEX159), 0.1μ                                                        Ci/ml                                                Mouse    .sup.125 I goat anti-mouse                                                                      (See primary antibody)                               IgG      (NEN # NEX159, at 0.1                                                          μCi/ml                                                             Vaccinia Rabbit anti-vP410,                                                                              .sup.125 I Protein A (NEN                            proteins rabbit #W29 week 39,                                                                            #NEX-146), 0.1 μCi/ml                                      9/13/91, preabsorbed                                                           against Vero cells and                                                         diluted 1:100                                                         Vero     Rabbit anti-Vero cells,                                                                         .sup.25 I Protein A (NEN                             cell     obtained from B. #NEX-146), 0.1 μCi/ml                             proteins Meignier, PMsv,                                                                preabsorbed against                                                            ALVAC-infected CEF and                                                         diluted 1:100                                                         ______________________________________                                    

Immunoprecipitation/western blot assay. A combination IP/Western Blot was performed on Batch 1 gB utilizing the panel of monoclonal antibodies. Unlabeled crude and purified gB was subjected to immunoprecipitation followed by SDS-PAGE, the gel was blotted onto nitrocellulose, and gB-specific proteins detected with guinea pig anti-CMV gB (from Eva Gbnczbl), diluted 1:1000, and ¹²⁵ I Protein A (NEN #NEX146), 0.1μ Ci/ml.

Analysis of the purity of the gB product. Samples from each batch of gB were analyzed by electrophoretic separation on a 10% gel under reducing conditions, followed by staining with Coomassie Blue. The dried gel was scanned and analyzed by densitometry (PDI, Inc., Huntington Station, N.Y., Quantity One densitometer program).

Immunoprecipitation assay comparing expression of CMV gB by three vaccinia COPAK recombinants. To choose a suitable recombinant, cell substrate and antibody for production and immunoaffinity purification of CMV gB, COPAK recombinants expressing 3 different forms of gB were compared by immunoprecipitation assay using guinea pig antig-gB and a panel of monoclonal antibodies. Recombinants vP1126, vP1128, and vP1145 elicit CMV neutralizing antibodies in mice and therefore express gB in an immunogenic form (Table 23). All of the CMV gB antibodies tested produced similar IP results. A representative assay, with guinea pig serum using both medium and cell fractions from HeLa and Vero cell infections, is shown in FIGS. 52A to D. As expected, CMV gB specific material was precipitated from both the cell and medium fractions of vP1128 and vP1145 infected cells, but in only the cell fraction with vP1126 infected cells. The apparent molecular weights of the gB specific bands correspond to previously published results (Britt and Auger, 1986; Britt and Vugler, 1989; Reis et.al., 1993). The cell fractions of all three CMV gB recombinants contained a major band of apparent molecular weight 130-140 kDa, consistent with the apparent molecular weight of the glycosylated uncleaved gB precursor. Less intense protein species with apparent MW of 110 kDa and 55 kDa were observed in the cell fractions and are consistent with the proteolytically processed mature protein species. The N-terminal product was previously reported to be 90-110 kDa and the C-terminal product 55-58 kDa (Britt and Auger, 1986). In HeLa cells a protein species with an apparent higher molecular mass (approximately 150 kDa) was also present (e.g., FIG. 52D, lane 4). This species probably also represents an uncleaved precursor form that is more highly glcosylated. In the medium fractions three gB bands were precipitated from vP1128 and vP1145 infected cells, representing the uncleaved precursor, and N-terminal and C-terminal processed polypeptides. By densitometric analysis, there was more gB-specific material precipitated from the medium fractions of Vero cells compared to HeLa, with recombinant vP1145 producing more gB-specific material than vP1128. This difference may be explained by the observation that more vaccinia E3L was precipitated from the cell fraction of vP1145 than the vP1128 cell fraction, indicating an overall higher level of vaccinia expression in this sample (FIGS. 53A and B). With vP1145, there was more gB specific material precipitated from the medium fraction than from the cell fraction in both HeLa and Vero cells (compare FIG. 52 A,B vs. C,D).

The three different sizes of gB precipitated from the medium of HeLa infected cells appear to have higher molecular weights than the three species produced in Vero cells (compare FIG. 52A vs. 52B). These differences may be due to different levels of glycosylation in HeLa cells compared to Vero, but this hypothesis was not examined further. To determine if the higher molecular weight gB-specific proteins would also be produced by another human cell line, MRC-5, a western blot assay was performed comparing the gB proteins in the medium of vP1145 infected HeLa, MRC-5, and Vero cells using monoclonal CH380 (FIG. 54). The result shows that the two gB bands detectable in this assay, gB precursor (approx. 140 kDa) and C terminal processing fragment (55-58 kDa), had apparently higher molecular weights in HeLa and MRC-5 than in VERO cells. The N-terminal processing fragment is not detectable by western blot using either monoclonal CH380 or the guinea pig anti-CMV gB serum.

MAb CH380 was chosen for use in immunoaffinity purification of gB, since a large quantity was readily available and no apparent differences were seen in the gB-specific proteins detected by the five different monoclonals in the IP assay (FIG. 55). Based on the IP analysis and the considerations that purification of secreted gB from the medium of infected cells eliminates the need to solubilize gB from cell membranes and purify it from cellular proteins, purification of CMV gB was initiated using the medium fraction of vP1145-infected Vero cells. Infection was done in serum-free medium, further reducing contaminating proteins in the crude material.

Purification of CMV gB. Fifteen separate immunoaffinity chromatography column runs, yielding a total of 3.1 mg of gB, are summarized in Table 24. Some of the material was used for further assays and the remainder was pooled in 5 separate batches of purified product, totaling 2.6 mg (Table 25). Column runs 7, 8, 10, and 11 were sequential runs in the same column. The bed material from columns 19A, 19B, 19C, 21A, 21B, and 21C were pooled to make the column used for runs 28, 29, and 32, from which the largest amount of gB was obtained. Table 24 lists the Crude gB material applied to each column in terms of the number of vP1145-infected Vero roller bottles (1×10⁸ cells per RB) from which the crude material was derived, and amount of total protein and gB-specific protein in the crude. Based on analysis of 8 samples, the total protein content of the crude preparations ranged from 1.2 to 3.7 mg /RB with a mean value of 2.4 mg/RB (24 μg per 10⁶ cells). Utilizing a slot blot assay with purified gB as standard, the amount of gB present in the crude material was measured for 7 of the preparations: values ranged from 50 to 350 μg/RB with a mean of 153 μg/RB (1.5 μg/10⁶ cells). Together these calculations indicate that the protein in the crude preparations consisted of approximately 6% gB. CMV gB yields ranged from 8 to 29 μg/RB with a mean of 20 μg/RB (0.2 μg/10⁶ cells) (Table 24). Approximately fifty roller bottles (1×10⁹ cells) were required to produce 1 mg of CMV gB.

The capacity of the immunoabsorbent gel for gB was not fully evaluated. The 4 ml bed material used for column runs 28, 29, and 32, was initially divided into 0.6 ml mini-columns (column runs 19A, 19B, 19C, 21A, 21B, and 21C) and varying amounts of crude gB were applied to each column to determine where saturation of binding would occur. Unfortunately, the quantity of gB in the crude material applied to the columns was overestimated, and saturation was not demonstrated. The highest binding result (from column 19C) was used as an estimate of column capacity (300 μg/ml bed material). The amount of gB eluted from the minicolumns represented 8 to 25% of the gB protein applied to the columns (Table 24). Therefore, if the capacity of the 4 ml column is at least 1.2 mg and 25% of the gB applied is recovered, it was estimated that 4.9 mg of crude gB (from approximately 33 RB) must be applied to the column to obtain 1.2 mg of purified gB. The result from column 28 is close to this estimate: material from 36 roller bottles was applied to the column #28, and 1 mg of gB was eluted.

The gB applied to the columns but not eluted as purified material has not been quantitatively accounted for. Since only 8-25% of the gB applied to the column was recovered as purified gB, the remainder of the gB must be present in flow-through fractions, wash fractions, eluted fractions not pooled with the product, or bound to the column. CMV gB could be detected by western blot in the flow-through fractions (e.g., FIG. 56, lane 6). However, when the amount of gB in the flow-through fractions was estimated by slot blot analysis, it did not account for more than 20% of the applied gB. The wash fractions have not been evaluated. The pooled fractions chosen for the final gB product were peak fractions only and therefore trace amounts of gB in adjacent fractions could account for some of the missing gB. For example, FIG. 57 shows sequential fractions eluted from column 8. Fractions 8.17-8.21 were pooled for the gB product, but trace amounts remained in fractions 8.16 and 8.22. Evidence exists also for the retention of gB in the immunoabsorbent gel. Gel material, taken from columns 11 and 19C after elution and washes, contains gB specific material detectable by western blot (FIG. 56, lanes 2 and 3). The amount of gB remaining on the column has not been quantitatively evaluated.

Reapplication of flow-through material to the column was attempted when flow-through material from column run #7 was applied to column #10 (Table 24). The amount of gB eluted from column 10 (4.5 μg) was only 4% of that obtained from column 7 (110 μg). It was not possible to evaluate this result since the capacity of the bed material for gB, and the amounts of gB applied to the column and present in the flow-through fractions were not known. Because of the poor yield, this approach was not used again.

Evaluation of purified qB. After pooling gB-containing eluted fractions, evaluation of purified gB consisted of 1) determination of total protein concentration, 2) SDS-PAGE analysis to identify gB specific and non-specific bands, and 3) confirmation of these bands with immunological reagents. Additionally, the purified gB was analyzed for degree of purity by densitometer scan, and for native conformation by ability to bind to a panel of CMV monoclonal antibodies.

Fractions containing CMV gB eluted from each column were analyzed initially by SDS-PAGE and silver staining, and gB fractions were identified and pooled for each run. A typical elution profile is shown in FIG. 57. A portion of the eluted gB was used for analysis, and the remainder of the material was combined into 5 separate batches (Table 25). Each batch was analyzed by SDS-PAGE on a 10% gel under reducing conditions and stained with Coomassie Blue (FIG. 58). The stained gel was scanned on a densitometer and the molecular weight and relative quantity of each band was calculated: a typical scan is shown in FIGS. 59, 59A and analysis of the 5 batches is summarized in Table 26. By SDS-PAGE analysis batches 1-5 appear very similar (FIG. 58). The two major bands, having apparent molecular weights 120-130 and 51-59 kDa, represent the precursor gB protein and the C-terminal processing fragment. The wide diffuse appearance of these bands is probably due to variable glycosylation of this normally heavily glycosylated protein. The identity of these bands as gB-specific is supported by results from western blot analysis with monoclonal CH380 (FIG. 60B). The bands of apparent molecular weight 77-100 kDa, which appear as doublets in batches 2-5 (FIG. 58), are the correct size for the gB N-terminal processing fragment, identified in the medium of vP11145-infected cells by IP analysis (FIGS. 52A and B). These bands could not be verified as gB-specific by either western blot analysis (FIG. 60B), or a combination immunoprecipitation-western blot assay (FIGS. 61A and B), but the possibility should not be ruled out since neither the guinea pig anti-gB serum nor monoclonal CH380 are able to detect N-terminal processing fragments by western blot. A contaminating protein of approximately 39-45 kDa is present in each batch at a level of 6-15% of total protein (FIG. 58 and Table 26). Two more possible gB protein bands, one of greater than 200 kDa and the other 30-35 kDa are present in every batch (FIGS. 58, 59, and 59A; Table 26). Evidence that the large (˜200 kDa) protein is gB is derived from western blot analysis with monoclonal CH380 which detects two proteins with molecular weights greater than 200 kDa (FIG. 60B, lanes 2 & 3). It is possible that the protein of approximately 30-35 kDa is also gB-specific (FIG. 58). In the IP analysis of medium of vP1145-infected cells, a protein of approximately 35 kDa was detected by 3 monoclonals (13-128, HCMV 34, and HCMV 37)(FIG. 55) and by the guinea pig serum (FIG. 52A and B). A protein of this size was described by Reis et al. (1993) as a degradation product of gB.

Assuming that contaminating proteins in the gB preparation would be derived from the cell substrate, the virus vector or the immunoabsorbent bed material, the preparation was probed for the presence of mouse IgG, Vero cell proteins, and vaccinia proteins; Proteins derived from Vero cells or mouse IgG could not be detected by western blot analysis (FIGS. 60A and 62A). However, contaminating vaccinia-specific proteins with molecular weights of approximately 35 and 20 kDa were detected in trace amounts (FIG. 62B, lane 5).

To determine if the eluted gB retained its native conformation, a combination immunoprecipitation/western blot assay was performed with a panel of monoclonals which included 3 neutralizing and one conformationally dependent antibody. Each monoclonal antibody precipitated the precursor and C-terminal fragment from purified gB (FIG. 61), suggesting that the gB eluted from the immunoaffinity column retained its native conformation .

In summary, the analysis of eluted gB in batches 1-5 demonstrates that the product contains at least two known gB-specific proteins, the precursor gB and C-terminal fragment, which together account for approximately 50% of the protein content (FIG. 58 and Table 26). Three other protein species, which account for 20-25% of total protein content (Table 26), could also be gB-specific although direct evidence has not been provided.

Immunogenicity of purified gB. The five CMV gB batches were pooled and the final concentration determined. Several amounts of purified gB were adjuvanted with either alum or QS21 and used to inoculate mice. Serum from the mice was evaluated for the presence of HCMV neutralizing antibody. Table 27 demonstrates that all of the amounts of purified gB tested with both adjuvants were able to elicit HCMV neutralizing antibody.

Purified gB was used in a prime/boost protocol in combination with ALVAC-gB (vCP139) in mice. Table 28 demonstrates that mice receiving ALVAC gB (vCP139) on day 0 and boosted on Day 29 with purified gB adjuvanted with QS21 or Alum developed higher levels of HCMV neutralizing antibody than mice receiving a second dose of ALVAC-gB (vCP1319).

                  TABLE 23                                                         ______________________________________                                         Induction of HCMV Neutralizing Antibody in Mice                                        Days After Immunization                                                Immunogen.sup.1                                                                          30            48     135                                             ______________________________________                                         vP1126    .sup. 16.sup.2                                                                               8      256                                             vP1128    16            8      106                                             vP1145    16            8      106                                             ______________________________________                                          .sup.1 Mice were immunized with 1 × 10.sup.8 PFU of recombinant          viruses (ip.) on day 0 and day 49.                                             .sup.2 HCMV Neutralizing titer                                           

                                      TABLE 24                                     __________________________________________________________________________     SUMMARY OF IMMUNOAFFINITY PURIFICATION COLUMNS                                                    CRUDE MATERIAL                                                                 APPLIED                                                     # VERO             TO COLUMN                                                   COLUMN                                                                              ROLLER               gB-specific                                                                          gB YIELD                                       RUN  BOTTLES.sup.a                                                                        COLUMN SIZE                                                                            Total Protein.sup.b                                                                   protein.sup.c                                                                        (% of applied)                                 __________________________________________________________________________     7    4     1 ml    13.3 mg                                                                               nd.sup.d                                                                             110 ug.sup.b                                   8    6     1 ml    14.4 mg                                                                               2.2 mg                                                                               84 μg.sup.b                                 10   Col 7 1 ml    nd     nd    4.8 ug.sup.b                                        flow thru                                                                 11   4     1 ml    nd     nd    100 ug.sup.b                                   13   1     0.3 ml  nd     nd    12 ug.sup.d                                    19A  1     0.6 ml  2.9 mg 240 μg                                                                            41 μg.sup.c                                                                 (17%)                                          19B  2     0.6 ml  5.8 mg 480 μg                                                                            93 μg.sup.c                                                                 (19%)                                          19C  3     0.6 ml  8.7 mg 720 μg                                                                            185 μg.sup.c                                                                (25%)                                          21A  3     0.6 ml  5.7 mg 300 μg                                                                            29 μg.sup.c                                                                 (8%)                                           21B  5     0.6 ml  9.5 mg 500 μg                                                                            120 μg.sup.c                                                                (13%)                                          21C  7     0.6 ml  13.3 mg                                                                               700 μg                                                                            150 μg.sup.c                                                                (19%)                                          23   3     6 ml    5.7 mg 300 μg                                                                            25 μg.sup.c                                                                 (8%)                                           28   36    4 ml    64.8 mg                                                                               nd    1000 μg.sup.b                               29   24    4 ml    30 mg  nd    480 μg.sup.b                                32   24    4 ml    nd     nd    700 μg.sup.b                                __________________________________________________________________________      .sup.a Cells density: 1 × 10.sup.8 cells per roller bottle               .sup.b Protein concentration determined by Pierce BCA assay                    .sup.c Estimated by slot blot analysis, using purified gB as standard          .sup.d Not determined                                                    

                  TABLE 25                                                         ______________________________________                                         CMV gB BATCHES                                                                        TOTAL                          COLUMN                                   BATCH #                                                                               gB       VOLUME   CONCENTRATION                                                                               RUN                                      ______________________________________                                         1      0.16 mg  0.55 ml  0.29 mg/ml   7                                                                              8                                                                              10                                                                             11                                                                             13                                       2      1.0 mg   1.0 ml   1.0 mg/ml    28                                       3      0.26 mg  0.5 ml   0.52 mg/ml   21A                                                                            21B                                                                            21C                                                                            23                                       4      0.48 mg  0.5 ml   0.96 mg/ml   29                                       5      0.7 mg   0.5 ml   1.4 mg/ml    32                                       ______________________________________                                    

                                      TABLE 26                                     __________________________________________________________________________     DENSITOMETRY ANALYSIS OF 5 BATCHES OF CMV gB                                   PROTEIN                                                                              APPARENT MOLECULAR WEIGHT (kDa).sup.a                                                              RELATIVE QUANTITY (%).sup.b                          BAND  B1  B2  B3  B4  B5  B1 B2 B3 B4 B5                                       __________________________________________________________________________     >200 kDa                                                                             222 208 221 225 217 10.6                                                                              6.7                                                                               7.5                                                                               8.3                                                                               7.4                                      (gB?) 192                 8                                                    Precursor gB                                                                         128 120 124 128 134 39 30 36.1                                                                              30 27.4                                     N fragment                                                                           83  94  99  101 100 9.6                                                                               3.6                                                                               3.2                                                                               4.5                                                                               3.5                                      (?)       77  84  88  89     9.7                                                                               6.3                                                                               6.6                                                                               6.3                                      C fragment                                                                           55  51  55.4                                                                               56.4                                                                               59  21 15.6                                                                              13.7                                                                              22.6                                                                              21                                       Unknown                                                                              42  39  42  44  45  6.1                                                                               12 15.4                                                                              14.3                                                                              15.8                                     contaminant                                                                    gB    32  30  35  35  37  4.3                                                                               9.7                                                                               11.3                                                                              8.6                                                                               10                                       degradation                                                                    product (?)                                                                    __________________________________________________________________________      .sup.a Calculated from densitometer scan using molecular weight markers a      standards (refer to FIG. 59, 59A)                                              .sup.b The density of each band is calculated from a 2 dimensional scan        line through the band: the average pixel OD across the sample width is         integrated under the curve to the baseline to obtain density (ODxem).          Relative quantity is the percentage of the total density of all bands in       the lane.                                                                

                  TABLE 27                                                         ______________________________________                                         HCMV Neutralizing Antibodies Elicited by purified                              gB protein in CBA Mice.sup.1                                                                            NT.sup.2                                                                             NT.sup.2                                                                             NT.sup.2                                                                             NT.sup.2                            Mouse  dose.sup.3                                                                             Adjuvant.sup.3                                                                           4w    6w    8w    9w                                  ______________________________________                                         201    2.5     Alum      32    256   256   256                                 203                      8     64    128   128                                 204                      8     12    16    16                                  206    5.0     Alum      48    512   192   192                                 207                      12    192   512   512                                 208                      16    192   192   192                                 209                      16    128   256   256                                 210                      8     128   256   256                                 211    10.0    Alum      32    256                                             213                      32    96    256   256                                 214                      32    256   256                                       216    20.0    Alum      64    128   128   128                                 217                      64    256   256   256                                 218                      32    128   512   256                                 219                      16    128   256   256                                 220                      32    192   512   256                                 222    2.5     QS21      8     192   512                                       223                      32    >4096 >4096 2048                                224                      16    1536                                            225                      64    1024  1024  1024                                226    5.0     QS21      64    >4096 1624  1024                                227                      96    >4096                                           228                      64    >4096 >4096 >4096                               229                      64    >256  >4096                                     230                      32    >4096 1536  2048                                231    10.0    QS21      64    2048  2048  >4096                               232                      96    1536  2048                                      233                      96    >4096                                           234                      64    2048  2048  1024                                236    20.0    QS21      128   3072                                            239                      96    >4096 >4096 >4096                               ______________________________________                                          .sup.1 Mice were inoculated S.C. at weeks 0 and 4.                             .sup.2 Sera were obtained at 4, 6, 8 or 9 weeks after priming.                 .sup.3 μg gB in either 15 μg QS21 or 25 μl Alum were used for         each inoculation.                                                        

                  TABLE 28                                                         ______________________________________                                         Summary Of Prime-Boost Experiment                                                    NT      antigen  NT    antigen NT    NT                                  Mice  Day 0   adj.     Day 29                                                                               adj.    Day 42                                                                               Day 56                              ______________________________________                                         381   4       ALV      32    gB + Alu                                                                               384   768                                 382   <4      ALV      8     gB + Alu                                                                               192   192                                 383   4       ALV      4     gB + Alu                                                                               192   256                                 384   <4      ALV      48    gB + Alu                                                                               512   512                                 385   4       ALV      16    gB + Alu                                                                               256   ND                                  397   4       ALV      8     gB + Alu                                                                               128   192                                 G.m. 4                 13.5          248   326                                 392   <4      ALV      <4    gB + QS 128   128                                 393   <4      ALV      4     gB + QS >1024 >1024                               394   <4      ALV      8     gB + QS >1024 >1024                               395   <4      ALV      16    gB + QS 512   384                                 396   <4      ALV      4     gB + QS 256   384                                 398   4       ALV      8     gB + QS >1624 >1024                               G.m. 4                 6.3           >512  >522                                373   4       ALV      16    ALV     128   96                                  376   4       ALV      4     ALV     8     12                                  378   8       ALV      4     ALV     8     4                                   379   4       ALV      8     ALV     128   128                                 380   4       ALV      16    ALV     64    64                                  399   4       ALV      4     ALV     96    192                                 400   <4      ALV      4     ALV     64    128                                 G.m. 4                 6.5           45.6  51.2                                ______________________________________                                          5 × 10.sup.5 TCD.sub.50 of ALVACgB (vCP139), 5 μg gB + Alu, 1         μg gB + Q521 were given, s.c.                                               G.m. = geometric mean                                                    

The results presented here demonstrate the ability of the NYVAC and ALVAC-HCMV recombinants and products therefrom to be employed in the compositions and utilities aforementioned, for instance, immunological, antigenic or vaccine compositions, or for use in preparing antigens or antibodies for assays, kits or tests, and, for example, as suitable for uses in vaccine or immunization strategies capable of preventing infection by HCMV; and, that the DNA of the recombinants is useful for probes or for preparing PCR primers.

Example 47 Expression of CMV genes in NYVAC-CMV6 and NYVAC-CMV5

Immunoprecipitation with monoclonal antibodies specific for gB, gH, pp65, pp150 and IE1-exon4 demonstrated the correct expression of these five genes by NYVAC-CMV6. FACScan analysis (Becton-Dickinson ) demonstrated surface expression of gH in vP1302B infected cells but not in cells infected with its parent (vP1251) indicating that a functional gL gene product is expressed in vP1302B.

Immunoprecipitation with monoclonal antibodies specific for gB, gH, pp65and pp150 demonstrated the correct expression of these four genes by NYVAC-CMV. FACScan analysis demonstrated surface expression of gH in vP1312 infected cells but not in cells infected with its parent (vP1262) indicating that a functional gL gene product is expressed in vP1312.

Example 48 Developing an ALVAC Donor Plasmid Containing the HCMV pp65 And pp150 Genes

Plasmid CMV65C6.2 was linearized with EcoRI, filled in with klenow and treated with alkaline phosphatase generating a 6.3 kb fragment. Plasmid 150.1 was digested with NheI, filled in with klenow and a 3.2 kb fragment (42K-pp150) isolated. Ligation of these two fragments yielded plasmid 150.8R1 in which transcription of pp65 and pp150 are in the same direction and pp150 is reversed from plasmid 150.8 in example 40. The DNA sequence of CMVpp65 and CMVpp150 plus additional flanking sequences in plasmid 150.8R1 are shown in FIGS. 63 A-C (SEQ ID NO:177).

Example 49 Construction of ALVAC-CMV4 (gB, gH, pp65, pp150)

Plasmid 150.8R1 was transfected into vCP233 infected CEF cells to generate ALVAC-CMV4 (vP1360).

Example 50 Expression of CMV Genes iN ALVAC-CMV4

Immunoprecipitation with monoclonal antibodies specific for gB, gH, pp65 and pp150 demonstrated the correct expression of all four genes by ALVAC-CMV4 (vP1360).

Example 51 Developing ALVAC Donor Plasmids Containing HCMV gL or gL Plus IE1-exon4

FIGS. 64A and B (SEQ ID NO:178) is the sequence of a 5.8 kd segment of canarypox DNA contained in plasmid pcPtk. The canarypox thymidine kinase gene (tk) is encoded within this segment initiating at nucleotide 4412 and terminating at nucleotide 4951. A tk (C7) insertion vector containing 2085bp upstream of C7, polylinker containing SmaI, NruI, EcoRI, XhoI and StuI sites, and 812bp downstream of C7 was derived in the following manner. A 3450bp PstI/NsiI fragment from pCPtk was cloned into the blunt ended Asp718/XbaI sites of pBS-SK+ generating plasmid pEU1. To delete the tk ORF and replace it with a polylinker, two PCR fragments were amplified from pCPtk using oligonucleotides RG578 (SEQ ID NO:179) (5'-GTACATAAGCTTTTTGCATG-3') plus RG581 (SEQ ID NO:180) (5'-TATGAATTCCTCGAGGGATCCAGGCCTTTTTTATTGACTAGTTAATCAGTCTAATATACG TACTAAATAC-3') and RG579 (SEQ ID NO:181) (5'-CTAATTTCGAATGTCCGACG-3') plus RG580 (SEQ ID NO:182) (5'-TTAGAATTCTCGCGACCCGGGTTTTTATAGCTAATTAGTACTTATTACAAATACTATAAT ATTTAG-3'). These fragments were purified, digested with HindIII/EcoRI or BstBI/EcoRI and ligated to pEU1 cut with HindIII/BstBI resulting in plasmid pC7.

The polylinker region in pC7 was modified in the following manner. pC7 was digested with EcoRI and StuI, purified and ligated to annealed oligonucleotides SDSYN154 (SEQ ID NO:183) (5'-AATTCGTCGACGGAT CCCTCGAGGGTACCGCATGC-3') and SDSYN155 (SEQ ID NO:184) (5'-GCATGCGGTACCCTCGAGGGATCCGTCGACG-3') generating plasmid pC7⁺.

Plasmid pC7⁺ was digested with BamHI and treated with alkaline phosphatase. Plasmid I4LH6CgL was digested with BamHI and BglII and a 968bp fragment (containing the H6 promoted gL gene ) isolated. Ligation of these two fragments generated plasmid C7gL in which transcription of gL is in the same direction as the deleted tk gene. The DNA sequence of HCMV gL plus additional flanking sequences in plasmid C7gL is shown in FIGS. 65A and B SEQ ID NO:189.

Plasmid C7gL was digested with BamHI and PspAI and treated with alkaline phosphatase. Plasmid I4LH6IEEX4 was digested with BamHI and PspAI and a 1363bp fragment (containing the H6 promoted IE1-exon4 gene) isolated. Ligation of these two fragments yielded plasmid C7gLIES2. The DNA sequence of HCMV gL and IE1-exon4 plus additional flanking sequences in plasmid C7gLIES2 is shown in FIGS. 66A and B SEQ ID NO190.

Example 52 Construction of ALVAC-CMV6 (gB, gH, gL, pp65, pp150, IE1-exon4 and ALVAC-CMV5 (gB, gH, gL, pp65, pp150)

Plasmid C7gLIES2 is transfected into vP1360 infected cells to generate ALVAC-CMV6 (gB, gH, gL, pp65, pp150, IE1-exon4).

Plasmid C7gL is transfected into vP1360 infected cells to generate ALVAC-CMV5 (gB, gH, gL, pp65, pp150).

Example 53 Cloning of HCMV gL and a gH Lacking its Transmembrane Region and Cytoplasmic Tail in NYVAC Donor Plasmid pSD553

The sequence of HCMV gH lacking its transmembrane region and cytoplasmic tail is presented in FIG. 67 (SEQ ID NO:188). Plasmid SPgH1 was used in PCR with oligonucleotides SPgHS1 (SEQ ID NO:185) (5'-CCGAAGCTTCTCGAGAT AAAAATCAACGACTGTCGGTAGCGTCCACGACGAC-3') and SPgH8 (SEQ ID NO:186) (5'-TCCACTCCATGCTAGT-3') to generate a 756bp fragment. This fragment was digested with NsiI and HindIII and a 275bp fragment isolated. Plasmid SPgH6 was digested with NsiI and HindIII and a 4779bp fragment isolated. Ligation of these two fragments yielded plasmid SPgH7 which contains the 42K promoted gH gene lacking its transmembrane region and cytoplasmic tail.

NYVAC insertion plasmid pSD553VC was digested with BamHI and treated with alkaline phosphatase. Plasmid I4LH6CgL was digested with BamHI and BglII and a 970 bp fragment (containing the H6 promoter and gL gene) isolated. Ligation of these two fragments generated plasmid COPAKgL-24.

Plasmid gH7 was digested with XhoI and ScaI and a 2239bp fragment isolated (containing the 42K promoter and truncated gH gene). Plasmid COPAKgL-24 was digested with XhoI, treated with alkaline phosphatase and ligated to the 2239bp fragment generating plasmid COPAKHL-15. The DNA sequence of gL and the truncated gH plus additional flanking DNA sequences in plasmid COPAKHL-15 is shown in FIGS. 68A and B (SEQ ID NO:187).

Example 54 Constructing a Poxvirus Recombinant Containing gL and gH Lacking its Transmembrane Region and Cytoplasmic Tail

Plasmid COPAKHL-15 was transfected into NYVAC infected CEF cells to generate the recombinant vP1399.

Example 55 Expression of gH by Recombinant vP1399

Immunoprecipitation with a monoclonal antibody specific for gH revealed the expression of a secreted gH protein of approximately 97 kDa by recombinant vP1399.

Having thus described in detail preferred embodiments of the present invention, it is to be understood that the invention defined by the appended claims is not to be limited by particular details set forth in the above description as many apparent variations thereof are possible without departing from the spirit or scope thereof.

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    __________________________________________________________________________     #             SEQUENCE LISTING                                                 - (1) GENERAL INFORMATION:                                                     -    (iii) NUMBER OF SEQUENCES: 190                                            - (2) INFORMATION FOR SEQ ID NO:1:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:1:                                  # 20               CGGG                                                        - (2) INFORMATION FOR SEQ ID NO:2:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 28 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:                                  #             28   GTTA ATTACATG                                               - (2) INFORMATION FOR SEQ ID NO:3:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 73 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3:                                  - AGCTTCCCGG GTAAGTAATA CGTCAAGGAG AAAACGAAAC GATCTGTAGT TA - #GCGGCCGC          60                                                                           #      73                                                                      - (2) INFORMATION FOR SEQ ID NO:4:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 69 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:                                  - ATTAGTTAAT TAGGCGGCCG CTAACTACAG ATCGTTTCGT TTTCTCCTTG AC - #GTATTACT          60                                                                           #         69                                                                   - (2) INFORMATION FOR SEQ ID NO:5:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:                                  # 20               CCGC                                                        - (2) INFORMATION FOR SEQ ID NO:6:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 22 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:                                  #                 22CCG TT                                                     - (2) INFORMATION FOR SEQ ID NO:7:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:                                  # 20               TAAT                                                        - (2) INFORMATION FOR SEQ ID NO:8:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 41 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:                                  #   41             GCTC CCCGGGCTCG AGGGATCCGT T                                - (2) INFORMATION FOR SEQ ID NO:9:                                             -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 39 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:                                  #    39            CCGG GGAGCTCAGA TCTAGTAAT                                   - (2) INFORMATION FOR SEQ ID NO:10:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 16 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:10:                                 #    16                                                                        - (2) INFORMATION FOR SEQ ID NO:11:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 12 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:11:                                 #       12                                                                     - (2) INFORMATION FOR SEQ ID NO:12:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 75 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:                                 - TATGAGTAAC TTAACTCTTT TGTTAATTAA AAGTATATTC AAAAAATAAG TT - #ATATAAAT          60                                                                           #    75                                                                        - (2) INFORMATION FOR SEQ ID NO:13:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 73 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:13:                                 - AACGAATTCA GATCTATTTA TATAACTTAT TTTTTGAATA TACTTTTAAT TA - #ACAAAAGA          60                                                                           #      73                                                                      - (2) INFORMATION FOR SEQ ID NO:14:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 49 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:14:                                 #               49TGTTA ACTTTATATA ACTTATTTTT TGAATATAC                        - (2) INFORMATION FOR SEQ ID NO:15:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 67 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15:                                 - ACACGAATGA TTTTCTAAAG TATTTGGAAA GTTTTATAGG TAGTTGATAG AA - #CAAAATAC          60                                                                           #          67                                                                  - (2) INFORMATION FOR SEQ ID NO:16:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 51 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:                                 #             51TATAAAA CTTTCCAAAT ACTTTAGAAA ATCATTCGTG T                     - (2) INFORMATION FOR SEQ ID NO:17:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 46 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:                                 #                 46TTT TATACTAAGA TCTCCCGGGC TGCAGC                           - (2) INFORMATION FOR SEQ ID NO:18:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 66 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18:                                 - GGCCGCTGCA GCCCGGGAGA TCTTAGTATA AAAAGTGATT TATTTTTACA AA - #ATTATGTA          60                                                                           #           66                                                                 - (2) INFORMATION FOR SEQ ID NO:19:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 50 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19:                                 #              50CACCAA TTTAGATCTT ACTCAAAATA TGTAACAATA                       - (2) INFORMATION FOR SEQ ID NO:20:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 44 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:                                 # 44               ACTC ATATTAATAA AAATAATATT TATT                             - (2) INFORMATION FOR SEQ ID NO:21:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 72 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:                                 - GATCCTGAGT ACTTTGTAAT ATAATGATAT ATATTTTCAC TTTATCTCAT TT - #GAGAATAA          60                                                                           #       72                                                                     - (2) INFORMATION FOR SEQ ID NO:22:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 72 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:                                 - AATTCCTAAG ATCTTTTTAT TCTCAAATGA GATAAAGTGA AAATATATAT CA - #TTATATTA          60                                                                           #       72                                                                     - (2) INFORMATION FOR SEQ ID NO:23:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 72 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:                                 - GATCCAGATC TCCCGGGAAA AAAATTATTT AACTTTTCAT TAATAGGGAT TT - #GACGTATG          60                                                                           #       72                                                                     - (2) INFORMATION FOR SEQ ID NO:24:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 72 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:24:                                 - AATTCCTAGT ACGCATCATA CGTCAAATCC CTATTAATGA AAAGTTAAAT AA - #TTTTTTTC          60                                                                           #       72                                                                     - (2) INFORMATION FOR SEQ ID NO:25:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 40 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:                                 #    40            TGCA GGGCGCCGGA TCCTTTTTCT                                  - (2) INFORMATION FOR SEQ ID NO:26:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 40 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:26:                                 #    40            GCCC TGCAGCTCGA GAGATCTCCC                                  - (2) INFORMATION FOR SEQ ID NO:27:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 3209 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:                                 - TGAATGTTAA ATGTTATACT TTGGATGAAG CTATAAATAT GCATTGGAAA AA - #TAATCCAT          60                                                                           - TTAAAGAAAG GATTCAAATA CTACAAAACC TAAGCGATAA TATGTTAACT AA - #GCTTATTC         120                                                                           - TTAACGACGC TTTAAATATA CACAAATAAA CATAATTTTT GTATAACCTA AC - #AAATAACT         180                                                                           - AAAACATAAA AATAATAAAA GGAAATGTAA TATCGTAATT ATTTTACTCA GG - #AATGGGGT         240                                                                           - TAAATATTTA TATCACGTGT ATATCTATAC TGTTATCGTA TACTCTTTAC AA - #TTACTATT         300                                                                           - ACGAATATGC AAGAGATAAT AAGATTACGT ATTTAAGAGA ATCTTGTCAT GA - #TAATTGGG         360                                                                           - TACGACATAG TGATAAATGC TATTTCGCAT CGTTACATAA AGTCAGTTGG AA - #AGATGGAT         420                                                                           - TTGACAGATG TAACTTAATA GGTGCAAAAA TGTTAAATAA CAGCATTCTA TC - #GGAAGATA         480                                                                           - GGATACCAGT TATATTATAC AAAAATCACT GGTTGGATAA AACAGATTCT GC - #AATATTCG         540                                                                           - TAAAAGATGA AGATTACTGC GAATTTGTAA ACTATGACAA TAAAAAGCCA TT - #TATCTCAA         600                                                                           - CGACATCGTG TAATTCTTCC ATGTTTTATG TATGTGTTTC AGATATTATG AG - #ATTACTAT         660                                                                           - AAACTTTTTG TATACTTATA TTCCGTAAAC TATATTAATC ATGAAGAAAA TG - #AAAAAGTA         720                                                                           - TAGAAGCTGT TCACGAGCGG TTGTTGAAAA CAACAAAATT ATACATTCAA GA - #TGGCTTAC         780                                                                           - ATGTACGTCT GTGAGGCTAT CATGGATAAT GACAATGCAT CTCTAAATAG GT - #TTTTGGAC         840                                                                           - AATGGATTCG ACCCTAACAC GGAATATGGT ACTCTACAAT CTCCTCTTGA AA - #TGGCTGTA         900                                                                           - ATGTTCAAGA ATACCGAGGC TATAAAAATC TTGATGAGGT ATGGAGCTAA AC - #CTGTAGTT         960                                                                           - ACTGAATGCA CAACTTCTTG TCTGCATGAT GCGGTGTTGA GAGACGACTA CA - #AAATAGTA        1020                                                                           - AAAGATCTGT TGAAGAATAA CTATGTAAAC AATGTTCTTT ACAGCGGAGG CT - #TTACTCCT        1080                                                                           - TTGTGTTTGG CAGCTTACCT TAACAAAGTT AATTTGGTTA AACTTCTATT GG - #CTCATTCG        1140                                                                           - GCGGATGTAG ATATTTCAAA CACGGATCGG TTAACTCCTC TACATATAGC CG - #TATCAAAT        1200                                                                           - AAAAATTTAA CAATGGTTAA ACTTCTATTG AACAAAGGTG CTGATACTGA CT - #TGCTGGAT        1260                                                                           - AACATGGGAC GTACTCCTTT AATGATCGCT GTACAATCTG GAAATATTGA AA - #TATGTAGC        1320                                                                           - ACACTACTTA AAAAAAATAA AATGTCCAGA ACTGGGAAAA ATTGATCTTG CC - #AGCTGTAA        1380                                                                           - TTCATGGTAG AAAAGAAGTG CTCAGGCTAC TTTTCAACAA AGGAGCAGAT GT - #AAACTACA        1440                                                                           - TCTTTGAAAG AAATGGAAAA TCATATACTG TTTTGGAATT GATTAAAGAA AG - #TTACTCTG        1500                                                                           - AGACACAAAA GAGGTAGCTG AAGTGGTACT CTCAAAATGC AGAACGATGA CT - #GCGAAGCA        1560                                                                           - AGAAGTAGAG AAATAACACT TTATGACTTT CTTAGTTGTA GAAAAGATAG AG - #ATATAATG        1620                                                                           - ATGGTCATAG ATAACTCTGA TATTGCAAGT AAATGCAATA ATAAGTTAGA TT - #TATTTAAA        1680                                                                           - AGGATAGTTA AAAATAGAAA AAAAGAGTTA ATTTGTAGGG TTAAAATAAT AC - #ATAAGATC        1740                                                                           - TTAAAATTTA TAAATACGCA TAATAATAAA AATAGATTAT ACTTATTACC TT - #CAGAGATA        1800                                                                           - AAATTTAAGA TATTTACTTA TTTAACTTAT AAAGATCTAA AATGCATAAT TT - #CTAAATAA        1860                                                                           - TGAAAAAAAA GTACATCATG AGCAACGCGT TAGTATATTT TACAATGGAG AT - #TAACGCTC        1920                                                                           - TATACCGTTC TATGTTTATT GATTCAGATG ATGTTTTAGA AAAGAAAGTT AT - #TGAATATG        1980                                                                           - AAAACTTTAA TGAAGATGAA GATGACGACG ATGATTATTG TTGTAAATCT GT - #TTTAGATG        2040                                                                           - AAGAAGATGA CGCGCTAAAG TATACTATGG TTACAAAGTA TAACTCTATA CT - #ACTAATGG        2100                                                                           - CGACTTCTGC AAGAAGGTAT AGTATAGTGA AAATGTTGTT AGATTATGAT TA - #TGAAAAAC        2160                                                                           - CAAATAAATC AGATCCATAT CTAAAGGTAT CTCCTTTGCA CATAATTTCA TC - #TATTCCTA        2220                                                                           - GTTTAGAATA CTTTTCATTA TATTTGTTTA CAGCTGAAGA CGAAAAAAAT AT - #ATCGATAA        2280                                                                           - TAGAAGATTA TGTTAACTCT GCTAATAAGA TGAAATTGAA TGAGTCTGTG AT - #AATAGCTA        2340                                                                           - TAATCAGAGA AGTTCTAAAA GGAAATAAAA ATCTAACTGA TCAGGATATA AA - #AACATTGG        2400                                                                           - CTGATGAAAT CAACAAGGAG GAACTGAATA TAGCTAAACT ATTGTTAGAT AG - #AGGGGCCA        2460                                                                           - AAGTAAATTA CAAGGATGTT TACGGTTCTT CAGCTCTCCA TAGAGCTGCT AT - #TGGTAGGA        2520                                                                           - AACAGGATAT GATAAAGCTG TTAATCGATC ATGGAGCTGA TGTAAACTCT TT - #AACTATTG        2580                                                                           - CTAAAGATAA TCTTATTAAA AAAAAATAAT ATCACGTTTA GTAATATTAA AA - #TATATTAA        2640                                                                           - TAACTCTATT ACTAATAACT CCAGTGGATA TGAACATAAT ACGAAGTTTA TA - #CATTCTCA        2700                                                                           - TCAAAATCTT ATTGACATCA AGTTAGATTG TGAAAATGAG ATTATGAAAT TA - #AGGAATAC        2760                                                                           - AAAAATAGGA TGTAAGAACT TACTAGAATG TTTTATCAAT AATGATATGA AT - #ACAGTATC        2820                                                                           - TAGGGCTATA AACAATGAAA CGATTAAAAA TTATAAAAAT CATTTCCCTA TA - #TATAATAC        2880                                                                           - GCTCATAGAA AAATTCATTT CTGAAAGTAT ACTAAGACAC GAATTATTGG AT - #GGAGTTAT        2940                                                                           - AAATTCTTTT CAAGGATTCA ATAATAAATT GCCTTACGAG ATTCAGTACA TT - #ATACTGGA        3000                                                                           - GAATCTTAAT AACCATGAAC TAAAAAAAAT TTTAGATAAT ATACATTAAA AA - #GGTAAATA        3060                                                                           - GATCATCTGT TATTATAAGC AAAGATGCTT GTTGCCAATA ATATACAACA GG - #TATTTGTT        3120                                                                           - TTTATTTTTA ACTACATATT TGATGTTCAT TCTCTTTATA TAGTATACAC AG - #AAAATTCA        3180                                                                           #          3209    TCTA GTTATCTAG                                              - (2) INFORMATION FOR SEQ ID NO:28:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 29 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:                                 #            29    AGCT AGCTAGTTT                                              - (2) INFORMATION FOR SEQ ID NO:29:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 46 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:                                 #                 46GGG AATTCTAGCT AGCTAGTTTT TATAAA                           - (2) INFORMATION FOR SEQ ID NO:30:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 50 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:                                 #              50CTAGCT AGCTAGAATT CCCGGGAAGC TTTTGAGAGT                       - (2) INFORMATION FOR SEQ ID NO:31:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 71 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:                                 - CTGAAATTAT TTCATTATCG CGATATCCGT TAAGTTTGTA TCGTAATGGT TC - #CTCAGGCT          60                                                                           #       71                                                                     - (2) INFORMATION FOR SEQ ID NO:32:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 48 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32:                                 #                48TAAC GGATATCGCG ATAATGAAAT AATTTCAG                         - (2) INFORMATION FOR SEQ ID NO:33:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 73 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33:                                 - ACCCCTTCTG GTTTTTCCGT TGTGTTTTGG GAAATTCCCT ATTTACACGA TC - #CCAGACAA          60                                                                           #      73                                                                      - (2) INFORMATION FOR SEQ ID NO:34:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 51 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:                                 #             51TTGTCTG GGATCGTGTA AATAGGGAAT TTCCCAAAAC A                     - (2) INFORMATION FOR SEQ ID NO:35:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 45 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:                                 #45                AGGG GTACAAACAG GAGAGCCTGA GGAAC                            - (2) INFORMATION FOR SEQ ID NO:36:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 11 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:                                 #       11                                                                     - (2) INFORMATION FOR SEQ ID NO:37:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 2724 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:                                 - ATGGAATCCA GGATCTGGTG CCTGGTAGTC TGCGTTAACT TGTGTATCGT CT - #GTCTGGGT          60                                                                           - GCTGCGGTTT CCTCATCTTC TACTCGTGGA ACTTCTGCTA CTCACAGTCA CC - #ATTCCTCT         120                                                                           - CATACGACGT CTGCTGCTCA TTCTCGATCC GGTTCAGTCT CTCAACGCGT AA - #CTTCTTCC         180                                                                           - CAAACGGTCA GCCATGGTGT TAACGAGACC ATCTACAACA CTACCCTCAA GT - #ACGGAGAT         240                                                                           - GTGGTGGGGG TCAACACCAC CAAGTACCCC TATCGCGTGT GTTCTATGGC AC - #AGGGTACG         300                                                                           - GATCTTATTC GCTTTGAACG TAATATCGTC TGCACCTCGA TGAAGCCCAT CA - #ATGAAGAC         360                                                                           - CTGGACGAGG GCATCATGGT GGTCTACAAA CGCAACATCG TCGCGCACAC CT - #TTAAGGTA         420                                                                           - CGAGTCTACC AGAAGGTTTT GACGTTTCGT CGTAGCTACG CTTACATCCA CA - #CCACTTAT         480                                                                           - CTGCTGGGCA GCAACACGGA ATACGTGGCG CCTCCTATGT GGGAGATTCA TC - #ATATCAAC         540                                                                           - AGTCACAGTC AGTGCTACAG TTCCTACAGC CGCGTTATAG CAGGCACGGT TT - #TCGTGGCT         600                                                                           - TATCATAGGG ACAGCTATGA AAACAAAACC ATGCAATTAA TGCCCGACGA TT - #ATTCCAAC         660                                                                           - ACCCACAGTA CCCGTTACGT GACGGTCAAG GATCAATGGC ACAGCCGCGG CA - #GCACCTGG         720                                                                           - CTCTATCGTG AGACCTGTAA TCTGAATTGT ATGGTGACCA TCACTACTGC GC - #GCTCCAAG         780                                                                           - TATCCCTATC ATTTTTTCGC AACTTCCACG GGTGATGTGG TTGACATTTC TC - #CTTTCTAC         840                                                                           - AACGGAACTA ATCGCAATGC CAGCTATTTT GGAGAAAACG CCGACAAGTT TT - #TCATTTTT         900                                                                           - CCGAACTACA CTATCGTCTC CGACTTTGAA AGACCGAATT CTGCGTTAGA GA - #CCCACAGG         960                                                                           - TTGGTGGCTT TTCTTGAACG TGCGGACTCA GTGATCTCCT GGGATATACA GG - #ACGAGAAG        1020                                                                           - AATGTTACTT GTCAACTCAC TTTCTGGGAA GCCTCGGAAC GCACCATTCG TT - #CCGAAGCC        1080                                                                           - GAGGACTCGT ATCACTTTTC TTCTGCCAAA ATGACCGCCA CTTTCTTATC TA - #AGAAGCAA        1140                                                                           - GAGGTGAACA TGTCCGACTC TGCGCTGGAC TGTGTACGTG ATGAGGCCAT AA - #ATAAGTTA        1200                                                                           - CAGCAGATTT TCAATACTTC ATACAATCAA ACATATGAAA AATATGGAAA CG - #TGTCCGTC        1260                                                                           - TTTGAAACCA CTGGTGGTTT GGTGGTGTTC TGGCAAGGTA TCAAGCAAAA AT - #CTCTGGTG        1320                                                                           - GAACTCGAAC GTTTGGCCAA CCGCTCCAGT CTGAATCTTA CTCATAATAG AA - #CCAAAAGA        1380                                                                           - AGTACAGATG GCAACAATGC AACTCATTTA TCCAACATGG AGTCGGTGCA CA - #ATCTGGTC        1440                                                                           - TACGCCCAGC TGCAGTTCAC CTATGACACG TTGCGCGGTT ACATCAACCG GG - #CGCTGGCC        1500                                                                           - GAAATCGCAG AAGCCTGGTG TGTGGATCAA CGGCGCACCC TAGAGGTCTT CA - #AGGAACTT        1560                                                                           - AGCAAGATCA ACCCGTCAGC TATTCTCTCG GCCATCTACA ACAAACCGAT TG - #CCGCGCGT        1620                                                                           - TTCATGGGTG ATGTCCTGGG TCTGGCCAGC TGCGTGACCA TTAACCAAAC CA - #GCGTCAAG        1680                                                                           - GTGCTGCGTG ATATGAATGT GAAGGAATCG CCAGGACGCT GCTACTCACG AC - #CAGTGGTC        1740                                                                           - ATCTTTAATT TCGCCAACAG CTCGTACGTG CAGTACGGTC AACTGGGCGA GG - #ATAACGAA        1800                                                                           - ATCCTGTTGG GCAACCACCG CACTGAGGAA TGTCAGCTTC CCAGCCTCAA GA - #TCTTCATC        1860                                                                           - GCCGGCAACT CGGCCTACGA GTACGTGGAC TACCTCTTCA AACGCATGAT TG - #ACCTCAGC        1920                                                                           - AGCATCTCCA CCGTCGACAG CATGATCGCC CTAGACATCG ACCCGCTGGA AA - #ACACCGAC        1980                                                                           - TTCAGGGTAC TGGAACTTTA CTCGCAGAAA GAATTGCGTT CCAGCAACGT TT - #TTGATCTC        2040                                                                           - GAGGAGATCA TGCGCGAGTT CAATTCGTAT AAGCAGCGGG TAAAGTACGT GG - #AGGACAAG        2100                                                                           - GTAGTCGACC CGCTGCCGCC CTACCTCAAG GGTCTGGACG ACCTCATGAG CG - #GCCTGGGC        2160                                                                           - GCCGCGGGAA AGGCCGTTGG CGTAGCCATT GGGGCCGTGG GTGGCGCGGT GG - #CCTCCGTG        2220                                                                           - GTCGAAGGCG TTGCCACCTT CCTCAAAAAC CCCTTCGGAG CCTTCACCAT CA - #TCCTCGTG        2280                                                                           - GCCATAGCCG TCGTCATTAT CATTTATTTG ATCTATATCC GACAGCGGCG TC - #TCTGCATG        2340                                                                           - CAGCCGCTGC AGAACCTCTT TCCCTATCTG GTGTCCGCCG ACGGGACCAC CG - #TGACGTCG        2400                                                                           - GGCAACACCA AAGACACGTC GTTACAGGCT CCGCCTTCCT ACGAGGAAAG TG - #TTTATAAT        2460                                                                           - TCTGGTCGCA AAGGACCGGG ACCACCGTCG TCTGATGCAT CCACGGCGGC TC - #CGCCTTAC        2520                                                                           - ACCAACGAGC AGGCTTACCA GATGCTTCTG GCCCTGGTCC GTCTGGACGC AG - #AGCAGCGA        2580                                                                           - GCGCACGAGA ACGGTACAGA TTCTTTGGAC GGACAGACTG GCACGCAGGA CA - #AGGGACAG        2640                                                                           - AAGCCCAACC TGCTAGACCG ACTGCGACAC CGCAAAAACG GCTACCGACA CT - #TGAAAGAC        2700                                                                           #              2724ACGT CTGA                                                   - (2) INFORMATION FOR SEQ ID NO:38:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 4260 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:                                 - AAGCTTTTGC GATCAATAAA TGGATCACAA CCAGTATCTC TTAACGATGT TC - #TTCGCAGA          60                                                                           - TGATGATTCA TTTTTTAAGT ATTTGGCTAG TCAAGATGAT GAATCTTCAT TA - #TCTGATAT         120                                                                           - ATTGCAAATC ACTCAATATC TAGACTTTCT GTTATTATTA TTGATCCAAT CA - #AAAAATAA         180                                                                           - ATTAGAAGCC GTGGGTCATT GTTATGAATC TCTTTCAGAG GAATACAGAC AA - #TTGACAAA         240                                                                           - ATTCACAGAC TCTCAAGATT TTAAAAAACT GTTTAACAAG GTCCCTATTG TT - #ACAGATGG         300                                                                           - AAGGGTCAAA CTTAATAAAG GATATTTGTT CGACTTTGTG ATTAGTTTGA TG - #CGATTCAA         360                                                                           - AAAAGAATCC TCTCTAGCTA CCACCGCAAT AGATCCTATT AGATACATAG AT - #CCTCGTCG         420                                                                           - CGATATCGCA TTTTCTAACG TGATGGATAT ATTAAAGTCG AATAAAGTGA AC - #AATAATTA         480                                                                           - ATTCTTTATT GTCATCATGT AATTAACTAG CTACCCGGGA GATCTCTCGA GC - #TGCAGAAG         540                                                                           - CTTATAAAAA TCACAAGTCT CTGTCACTTT TTTTGTCTAG TTTTTTTTTC TC - #CTCTTGGT         600                                                                           - TCAGACGTTC TCTTCTTCGT CGGAGTCTTT CAAGTGTCGG TAGCCGTTTT TG - #CGGTGTCG         660                                                                           - CAGTCGGTCT AGCAGGTTGG GCTTCTGTCC CTTGTCCTGC GTGCCAGTCT GT - #CCGTCCAA         720                                                                           - AGAATCTGTA CCGTTCTCGT GCGCTCGCTG CTCTGCGTCC AGACGGACCA GG - #GCCAGAAG         780                                                                           - CATCTGGTAA GCCTGCTCGT TGGTGTAAGG CGGAGCCGCC GTGGATGCAT CA - #GACGACGG         840                                                                           - TGGTCCCGGT CCTTTGCGAC CAGAATTATA AACACTTTCC TCGTAGGAAG GC - #GGAGCCTG         900                                                                           - TAACGACGTG TCTTTGGTGT TGCCCGACGT CACGGTGGTC CCGTCGGCGG AC - #ACCAGATA         960                                                                           - GGGAAAGAGG TTCTGCAGCG GCTGCATGCA GAGACGCCGC TGTCGAGTAT AG - #ATCAAATA        1020                                                                           - AATGATAATG ACGACGGCTA TGGCCACGAG GATGATGGTG AAGGCTCCGA AG - #GGGTTTTT        1080                                                                           - GAGGAAGGTG GCAACGCCTT CGACCACGGA GGCCACCGCG CCACCCACGG CC - #CCAATGGC        1140                                                                           - TACGCCAACG GCCTTTCCCG CGGCGCCCAG GCCGCTCATG AGGTCGTCCA GA - #CCCTTGAG        1200                                                                           - GTAGGGCGGC AGCGGGTCGA CTACCTTGTC CTCCACGTAC TTTACCCGCT GC - #TTATACGA        1260                                                                           - ATTGAACTCG CGCATGATCT CCTCGAGATC AAAAACGTTG CTGGAACGCA AT - #TCTTTCTG        1320                                                                           - CGAGTAAAGT TCCAGTACCC TGAAGTCGGT GTTTTCCAGC GGGTCGATGT CT - #AGGGCGAT        1380                                                                           - CATGCTGTCG ACGGTGGAGA TGCTGCTGAG GTCAATCATG CGTTTGAAGA GG - #TAGTCCAC        1440                                                                           - GTACTCGTAG GCCGAGTTGC CGGCGATGAA GATCTTGAGG CTGGGAAGCT GA - #CATTCCTC        1500                                                                           - AGTGCGGTGG TTGCCCAACA GGATTTCGTT ATCCTCGCCC AGTTGACCGT AC - #TGCACGTA        1560                                                                           - CGAGCTGTTG GCGAAATTAA AGATGACCAC TGGTCGTGAG TAGCAGCGTC CT - #GGCGATTC        1620                                                                           - CTTCACATTC ATATCACGCA GCACCTTGAC GCTGGTTTGG TTAATGGTCA CG - #CAGCTGGC        1680                                                                           - CAGACCCAGG ACATCACCCA TGAAACGCGC GGCAATCGGT TTGTTGTAGA TG - #GCCGAGAG        1740                                                                           - AATAGCTGAC GGGTTGATCT TGCTAAGTTC CTTGAAGACC TCTAGGGTGC GC - #CGTTGATC        1800                                                                           - CACACACCAG GCTTCTGCGA TTTCGGCCAG CGCCCGGTTG ATGTAACCGC GC - #AACGTGTC        1860                                                                           - ATAGGTGAAC TGCAGCTGGG CGTAGACCAG ATTGTGCACC GACTCCATGT TG - #GATAAATG        1920                                                                           - AGTTGCATTG TTGCCATCTG TACTTCTTTT GGTTCTATTA TGAGTAAGAT TC - #AGACTGGA        1980                                                                           - GCGGTTGGCC AAACGTTCGA GTTCCACCAG AGATTTTTGC TTGATACCTT GC - #CAGAACAC        2040                                                                           - CACCAAACCA CCAGTGGTTT CAAAGACGGA CACGTTTCCA TATTTTTCAT AT - #GTTTGATT        2100                                                                           - GTATGAAGTA TTGAAAATCT GCTGTAACTT ATTTATGGCC TCATCACGTA CA - #CAGTCCAG        2160                                                                           - CGCAGAGTCG GACATGTTCA CCTCTTGCTT CTTAGATAAG AAAGTGGCGG TC - #ATTTTGGC        2220                                                                           - AGAAGAAAAG TGATACGAGT CCTCGGCTTC GGAACGAATG GTGCGTTCCG AG - #GCTTCCCA        2280                                                                           - GAAAGTGAGT TGACAAGTAA CATTCTTCTC GTCCTGTATA TCCCAGGAGA TC - #ACTGAGTC        2340                                                                           - CGCACGTTCA AGAAAAGCCA CCAACCTGTG GGTCTCTAAC GCAGAATTCG GT - #CTTTCAAA        2400                                                                           - GTCGGAGACG ATAGTGTAGT TCGGAAAAAT GAAAAACTTG TCGGCGTTTT CT - #CCAAAATA        2460                                                                           - GCTGGCATTG CGATTAGTTC CGTTGTAGAA AGGAGAAATG TCAACCACAT CA - #CCCGTGGA        2520                                                                           - AGTTGCGAAA AAATGATAGG GATACTTGGA GCGCGCAGTA GTGATGGTCA CC - #ATACAATT        2580                                                                           - CAGATTACAG GTCTCACGAT AGAGCCAGGT GCTGCCGCGG CTGTGCCATT GA - #TCCTTGAC        2640                                                                           - CGTCACGTAA CGGGTACTGT GGGTGTTGGA ATAATCGTCG GGCATTAATT GC - #ATGGTTTT        2700                                                                           - GTTTTCATAG CTGTCCCTAT GATAAGCCAC GAAAACCGTG CCTGCTATAA CG - #CGGCTGTA        2760                                                                           - GGAACTGTAG CACTGACTGT GACTGTTGAT ATGATGAATC TCCCACATAG GA - #GGCGCCAC        2820                                                                           - GTATTCCGTG TTGCTGCCCA GCAGATAAGT GGTGTGGATG TAAGCGTAGC TA - #CGACGAAA        2880                                                                           - CGTCAAAACC TTCTGGTAGA CTCGTACCTT AAAGGTGTGC GCGACGATGT TG - #CGTTTGTA        2940                                                                           - GACCACCATG ATGCCCTCGT CCAGGTCTTC ATTGATGGGC TTCATCGAGG TG - #CAGACGAT        3000                                                                           - ATTACGTTCA AAGCGAATAA GATCCGTACC CTGAGCCATA GAACACACGC GA - #TAGGGGTA        3060                                                                           - CTTGGTGGTG TTGACCCCCA CCACATCTCC GTACTTGAGG GTAGTGTTGT AG - #ATGGTCTC        3120                                                                           - GTTAACACCA TGGCTGACCG TTTGGGAAGA AGTTACGCGT TGAGAGACTG AA - #CCGGATCG        3180                                                                           - AGAATGAGCA GCAGACGTCG TATGAGAGGA ATGGTGACTG TGAGTAGCAG AA - #GTTCCACG        3240                                                                           - AGTAGAAGAT GAGGAAACCG CAGCACCCAG ACAGACGATA CACAAGTTAA CG - #CAGACTAC        3300                                                                           - CAGGCACCAG ATCCTGGATT CCATTACGAT ACAAACTTAA CGGATATCGC GA - #TAATGAAA        3360                                                                           - TAATTTATGA TTATTTCTCG CTTTCAATTT AACACAACCC TCAAGAACCT TT - #GTATTTAT        3420                                                                           - TTTCACTTTT AAGTATAGAA TAAAGAAGCT TGCATGCCAC GCGTCTCGAG GG - #CCCCTGCA        3480                                                                           - GGTCGACTCT AGAGGATCCT GATCCTTTTT CTGGGTAAGT AATACGTCAA GG - #AGAAAACG        3540                                                                           - AAACGATCTG TAGTTAGCGG CCGCCTAATT AACTAATATT ATATTTTTTA TC - #TAAAAAAC        3600                                                                           - TAAAAATAAA CATTGATTAA ATTTTAATAT AATACTTAAA AATGGATGTT GT - #GTCGTTAG        3660                                                                           - ATAAACCGTT TATGTATTTT GAGGAAATTG ATAATGAGTT AGATTACGAA CC - #AGAAAGTG        3720                                                                           - CAAATGAGGT CGCAAAAAAA CTGCCGTATC AAGGACAGTT AAAACTATTA CT - #AGGAGAAT        3780                                                                           - TATTTTTTCT TAGTAAGTTA CAGCGACACG GTATATTAGA TGGTGCCACC GT - #AGTGTATA        3840                                                                           - TAGGATCGGC TCCTGGTACA CATATACGTT ATTTGAGAGA TCATTTCTAT AA - #TTTAGGAA        3900                                                                           - TGATTATCAA ATGGATGCTA ATTGACGGAC GCCATCATGA TCCTATTTTA AA - #TGGATTGC        3960                                                                           - GTGATGTGAC TCTAGTGACT CGGTTCGTTG ATGAGGAATA TCTACGATCC AT - #CAAAAAAC        4020                                                                           - AACTGCATCC TTCTAAGATT ATTTTAATTT CTGATGTGAG ATCCAAACGA GG - #AGGAAATG        4080                                                                           - AACCTAGTAC GGCGGATTTA CTAAGTAATT ACGCTCTACA AAATGTCATG AT - #TAGTATTT        4140                                                                           - TAAACCCCGT GGCGTCTAGT CTTAAATGGA GATGCCCGTT TCCAGATCAA TG - #GATCAAGG        4200                                                                           - ACTTTTATAT CCCACACGGT AATAAAATGT TACAACCTTT TGCTCCTTCA TA - #TTCAGCTG        4260                                                                           - (2) INFORMATION FOR SEQ ID NO:39:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 7351 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:                                 - AGATATTTGT TAGCTTCTGC CGGAGATACC GTGAAAATCT ATTTTCTGGA AG - #GAAAGGGA          60                                                                           - GGTCTTATCT ATTCTGTCAG CAGAGTAGGT TCCTCTAATG ACGAAGACAA TA - #GTGAATAC         120                                                                           - TTGCATGAAG GTCACTGTGT AGAGTTCAAA ACTGATCATC AGTGTTTGAT AA - #CTCTAGCG         180                                                                           - TGTACGAGTC CTTCTAACAC TGTGGTTTAT TGGCTGGAAT AAAAGGATAA AG - #ACACCTAT         240                                                                           - ACTGATTCAT TTTCATCTGT CAACGTTTCT CTAAGAGATT CATAGGTATT AT - #TATTACAT         300                                                                           - CGATCTAGAA GTCTAATAAC TGCTAAGTAT ATTATTGGAT TTAACGCGCT AT - #AAACGCAT         360                                                                           - CCAAAACCTA CAAATATAGG AGAAGCTTCT CTTATGAAAC TTCTTAAAGC TT - #TACTCTTA         420                                                                           - CTATTACTAC TCAAAAGAGA TATTACATTA ATTATGTGAT GAGGCATCCA AC - #ATATAAAG         480                                                                           - AAGACTAAAG CTGTAGAAGC TGTTATGAAG AATATCTTAT CAGATATATT AG - #ATGCATTG         540                                                                           - TTAGTTCTGT AGATCAGTAA CGTATAGCAT ACGAGTATAA TTATCGTAGG TA - #GTAGGTAT         600                                                                           - CCTAAAATAA ATCTGATACA GATAATAACT TTGTAAATCA ATTCAGCAAT TT - #CTCTATTA         660                                                                           - TCATGATAAT GATTAATACA CAGCGTGTCG TTATTTTTTG TTACGATAGT AT - #TTCTAAAG         720                                                                           - TAAAGAGCAG GAATCCCTAG TATAATAGAA ATAATCCATA TGAAAAATAT AG - #TAATGTAC         780                                                                           - ATATTTCTAA TGTTAACATA TTTATAGGTA AATCCAGGAA GGGTAATTTT TA - #CATATCTA         840                                                                           - TATACGCTTA TTACAGTTAT TAAAAATATA CTTGCAAACA TGTTAGAAGT AA - #AAAAGAAA         900                                                                           - GAACTAATTT TACAAAGTGC TTTACCAAAA TGCCAATGGA AATTACTTAG TA - #TGTATATA         960                                                                           - ATGTATAAAG GTATGAATAT CACAAACAGC AAATCGGCTA TTCCCAAGTT GA - #GAAACGGT        1020                                                                           - ATAATAGATA TATTTCTAGA TACCATTAAT AACCTTATAA GCTTGACGTT TC - #CTATAATG        1080                                                                           - CCTACTAAGA AAACTAGAAG ATACATACAT ACTAACGCCA TACGAGAGTA AC - #TACTCATC        1140                                                                           - GTATAACTAC TGTTGCTAAC AGTGACACTG ATGTTATAAC TCATCTTTGA TG - #TGGTATAA        1200                                                                           - ATGTATAATA ACTATATTAC ACTGGTATTT TATTTCAGTT ATATACTATA TA - #GTATTAAA        1260                                                                           - AATTATATTT GTATAATTAT ATTATTATAT TCAGTGTAGA AAGTAAAATA CT - #ATAAATAT        1320                                                                           - GTATCTCTTA TTTATAACTT ATTAGTAAAG TATGTACTAT TCAGTTATAT TG - #TTTTATAA        1380                                                                           - AAGCTAAATG CTACTAGATT GATATAAATG AATATGTAAT AAATTAGTAA TG - #TAGTATAC        1440                                                                           - TAATATTAAC TCACATTATG AATACTACTA ATCACGAAGA ATGCAGTAAA AC - #ATATGATA        1500                                                                           - CAAACATGTT AACAGTTTTA AAAGCCATTA GTAATAAACA GTACAATATA AT - #TAAGTCTT        1560                                                                           - TACTTAAAAA AGATATTAAT GTTAATAGAT TATTAACTAG TTATTCTAAC GA - #AATATATA        1620                                                                           - AACATTTAGA CATTACATTA TGTAATATAC TTATAGAACG TGCAGCAGAC AT - #AAACATTA        1680                                                                           - TAGATAAGAA CAATCGTACA CCGTTGTTTT ATGCGGTAAA GAATAATGAT TA - #TGATATGG        1740                                                                           - TTAAACTCCT ATTAAAAAAT GGCGCGAATG TAAATTTACA AGATAGTATA GG - #ATATTCAT        1800                                                                           - GTCTTCACAT CGCAGGTATA CATAATAGTA ACATAGAAAT AGTAGATGCA TT - #GATATCAT        1860                                                                           - ACAAACCAGA TTTAAACTCC CGCGATTGGG TAGGTAGAAC ACCGCTACAT AT - #CTTCGTGA        1920                                                                           - TAGAATCTAA CTTTGAAGCT GTGAAATTAT TATTAAAGTC AGGTGCATAT GT - #AGGTTTGA        1980                                                                           - AAGACAAATG TAAGCATTTT CCTATACACC ATTCTGTAAT GAAATTAGAT CA - #CTTAATAT        2040                                                                           - CAGGATTGTT ATTAAAATAT GGAGCAAATC CAAATACAAT TAACGGCAAT GG - #AAAAACAT        2100                                                                           - TATTAAGCAT TGCTGTAACA TCTAATAATA CACTACTGGT AGAACAGCTG CT - #GTTATATG        2160                                                                           - GAGCAGAAGT TAATAATGGT GGTTATGATG TTCCAGCTCC TATTATATCC GC - #TGTCAGTG        2220                                                                           - TTAACAATTA TGATATTGTT AAGATACTGA TACATAATGG TGCGAATATA AA - #TGTATCCA        2280                                                                           - CGGAAGATGG TAGAACGTCT TTACATACAG CTATGTTTTG GAATAACGCT AA - #AATAATAG        2340                                                                           - ATGAGTTGCT TAACTATGGA AGTGACATAA ACAGCGTAGA TACTTATGGT AG - #AACTCCGT        2400                                                                           - TATCTTGTTA TCGTAGCTTA AGTTATGATA TCGCTACTAA ACTAATATCA CG - #TATCATTA        2460                                                                           - TAACAGATGT CTATCGTGAA GCACCAGTAA ATATCAGCGG ATTTATAATT AA - #TTTAAAAA        2520                                                                           - CTATAGAAAA TAATGATATA TTCAAATTAA TTAAAGATGA TTGTATTAAA GA - #GATAAACA        2580                                                                           - TACTTAAAAG TATAACCCTT AATAAATTTC ATTCATCTGA CATATTTATA CG - #ATATAATA        2640                                                                           - CTGATATATG TTTATTAACG AGATTTATTC AACATCCAAA GATAATAGAA CT - #AACAAAAA        2700                                                                           - ACTCTACGCT TATAAATCTA TAGTCAACGA GAGAAAAATC AAAGCTACTT AC - #AGGTATTA        2760                                                                           - TCAAATAAAA AAAGTATTAA CTGTACTACC TTTTTCAGGA TATTTCTCTA TA - #TTGCCGTT        2820                                                                           - TGATGTGTTA GTATATATAC TTGAATTCAT CTATGATAAT AATATGTTGG TA - #CTTATGAG        2880                                                                           - AGCGTTATCA TTAAAATGAA ATAAAAAGCA TACAAGCTAT TGCTTCGCTA TC - #GTTACAAA        2940                                                                           - ATGGCAGGAA TTTTGTGTAA ACTAAGCCAC ATACTTGCCA ATGAAAAAAA TA - #GTAGAAAG        3000                                                                           - GATACTATTT TAATGGGATT AGATGTTAAG GTTCCTTGGG ATTATAGTAA CT - #GGGCATCT        3060                                                                           - GTTAACTTTT ACGACGTTAG GTTAGATACT GATGTTACAG ATTATAATAA TG - #TTACAATA        3120                                                                           - AAATACATGA CAGGATGTGA TATTTTTCCT CATATAACTC TTGGAATAGC AA - #ATATGGAT        3180                                                                           - CAATGTGATA GATTTGAAAA TTTCAAAAAG CAAATAACTG ATCAAGATTT AC - #AGACTATT        3240                                                                           - TCTATAGTCT GTAAAGAAGA GATGTGTTTT CCTCAGAGTA ACGCCTCTAA AC - #AGTTGGGA        3300                                                                           - GCGAAAGGAT GCGCTGTAGT TATGAAACTG GAGGTATCTG ATGAACTTAG AG - #CCCTAAGA        3360                                                                           - AATGTTCTGC TGAATGCGGT ACCCTGTTCG AAGGACGTGT TTGGTGATAT CA - #CAGTAGAT        3420                                                                           - AATCCGTGGA ATCCTCACAT AACAGTAGGA TATGTTAAGG AGGACGATGT CG - #AAAACAAG        3480                                                                           - AAACGCCTAA TGGAGTGCAT GTCCAAGTTT AGGGGGCAAG AAATACAAGT TC - #TAGGATGG        3540                                                                           - TATTAATAAG TATCTAAGTA TTTGGTATAA TTTATTAAAT AGTATAATTA TA - #ACAAATAG        3600                                                                           - ATAAATAACA TGATAACGGT TTTTATTAGA ATAAAATAGA GATAATATCA TA - #ATGATATA        3660                                                                           - TAATACTTCA TTACCAGAAA TGAGTAATGG AAGACTTATA AATGAACTGC AT - #AAAGCTAT        3720                                                                           - AAGGTATAGA GATATAAATT TAGTAAGGTA TATACTTAAA AAATGCAAAT AC - #AATAACGT        3780                                                                           - AAATATACTA TCAACGTCTT TGTATTTAGC CGTAAGTATT TCTGATATAG AA - #ATGGTAAA        3840                                                                           - ATTATTACTA GAACACGGTG CCGATATTTT AAAATGTAAA AATCCTCCTC TT - #CATAAAGC        3900                                                                           - TGCTAGTTTA GATAATACAG AAATTGCTAA ACTACTAATA GATTCTGGCG CT - #GACATAGA        3960                                                                           - ACAGATACAT TCTGGAAATA GTCCGTTATA TATTTCTGTA TATAGAAACA AT - #AAGTCATT        4020                                                                           - AACTAGATAT TTATTAAAAA AAGGTGTTAA TTGTAATAGA TTCTTTCTAA AT - #TATTACGA        4080                                                                           - TGTACTGTAT GATAAGATAT CTGATGATAT GTATAAAATA TTTATAGATT TT - #AATATTGA        4140                                                                           - TCTTAATATA CAAACTAGAA ATTTTGAAAC TCCGTTACAT TACGCTATAA AG - #TATAAGAA        4200                                                                           - TATAGATTTA ATTAGGATAT TGTTAGATAA TAGTATTAAA ATAGATAAAA GT - #TTATTTTT        4260                                                                           - GCATAAACAG TATCTCATAA AGGCACTTAA AAATAATTGT AGTTACGATA TA - #ATAGCGTT        4320                                                                           - ACTTATAAAT CACGGAGTGC CTATAAACGA ACAAGATGAT TTAGGTAAAA CC - #CCATTACA        4380                                                                           - TCATTCGGTA ATTAATAGAA GAAAAGATGT AACAGCACTT CTGTTAAATC TA - #GGAGCTGA        4440                                                                           - TATAAACGTA ATAGATGACT GTATGGGCAG TCCCTTACAT TACGCTGTTT CA - #CGTAACGA        4500                                                                           - TATCGAAACA ACAAAGACAC TTTTAGAAAG AGGATCTAAT GTTAATGTGG TT - #AATAATCA        4560                                                                           - TATAGATACC GTTCTAAATA TAGCTGTTGC ATCTAAAAAC AAAACTATAG TA - #AACTTATT        4620                                                                           - ACTGAAGTAC GGTACTGATA CAAAGTTGGT AGGATTAGAT AAACATGTTA TT - #CACATAGC        4680                                                                           - TATAGAAATG AAAGATATTA ATATACTGAA TGCGATCTTA TTATATGGTT GC - #TATGTAAA        4740                                                                           - CGTCTATAAT CATAAAGGTT TCACTCCTCT ATACATGGCA GTTAGTTCTA TG - #AAAACAGA        4800                                                                           - ATTTGTTAAA CTCTTACTTG ACCACGGTGC TTACGTAAAT GCTAAAGCTA AG - #TTATCTGG        4860                                                                           - AAATACTCCT TTACATAAAG CTATGTTATC TAATAGTTTT AATAATATAA AA - #TTACTTTT        4920                                                                           - ATCTTATAAC GCCGACTATA ATTCTCTAAA TAATCACGGT AATACGCCTC TA - #ACTTGTGT        4980                                                                           - TAGCTTTTTA GATGACAAGA TAGCTATTAT GATAATATCT AAAATGATGT TA - #GAAATATC        5040                                                                           - TAAAAATCCT GAAATAGCTA ATTCAGAAGG TTTTATAGTA AACATGGAAC AT - #ATAAACAG        5100                                                                           - TAATAAAAGA CTACTATCTA TAAAAGAATC ATGCGAAAAA GAACTAGATG TT - #ATAACACA        5160                                                                           - TATAAAGTTA AATTCTATAT ATTCTTTTAA TATCTTTCTT GACAATAACA TA - #GATCTTAT        5220                                                                           - GGTAAAGTTC GTAACTAATC CTAGAGTTAA TAAGATACCT GCATGTATAC GT - #ATATATAG        5280                                                                           - GGAATTAATA CGGAAAAATA AATCATTAGC TTTTCATAGA CATCAGCTAA TA - #GTTAAAGC        5340                                                                           - TGTAAAAGAG AGTAAGAATC TAGGAATAAT AGGTAGGTTA CCTATAGATA TC - #AAACATAT        5400                                                                           - AATAATGGAA CTATTAAGTA ATAATGATTT ACATTCTGTT ATCACCAGCT GT - #TGTAACCC        5460                                                                           - AGTAGTATAA AGTGATTTTA TTCAATTACG AAGATAAACA TTAAATTTGT TA - #ACAGATAT        5520                                                                           - GAGTTATGAG TATTTAACTA AAGTTACTTT AGGTACAAAT AAAATATTAT GT - #AATATAAT        5580                                                                           - AGAAAATTAT CTTGAGTCTT CATTTCCATC ACCGTCTAAA TTTATTATTA AA - #ACCTTATT        5640                                                                           - ATATAAGGCT GTTGAGTTTA GAAATGTAAA TGCTGTAAAA AAAATATTAC AG - #AATGATAT        5700                                                                           - TGAATATGTT AAAGTAGATA GTCATGGTGT CTCGCCTTTA CATATTATAG CT - #ATGCCTTC        5760                                                                           - AAATTTTTCT CTCATAGACG CTGACATGTA TTCAGAATTT AATGAAATTA GT - #AATAGACT        5820                                                                           - TCAAAAATCT AAAGATAGTA ACGAATTTCA ACGAGTTAGT CTACTAAGGA CA - #ATTATAGA        5880                                                                           - ATATGGTAAT GATAGTGATA TTAATAAGTG TCTAACATTA GTAAAAACGG AT - #ATACAGAG        5940                                                                           - TAACGAAGAG ATAGATATTA TAGATCTTTT GATAAATAAA GGAATAGATA TA - #AATATTAA        6000                                                                           - AGACGATTTA GGAAACACAG CTTTGCATTA CTCGTGTGAT TATGCTAAGG GA - #TCAAAGAT        6060                                                                           - AGCTAAAAAG TTACTAGATT GTGGAGCAGA TCCTAACATA GTTAATGATT TA - #GGTGTTAC        6120                                                                           - ACCACTAGCG TGTGCCGTTA ATACTTGCAA CGAGATACTA GTAGATATTC TG - #TTAAATAA        6180                                                                           - TGATGCGAAT CCTGATTCAT CTTCCTCATA TTTTTTAGGT ACTAATGTGT TA - #CATACAGC        6240                                                                           - CGTAGGTACC GGTAATATAG ATATTGTAAG ATCTTTACTT ACGGCTGGTG CC - #AATCCTAA        6300                                                                           - TGTAGGAGAT AAATCTGGAG TTACTCCTTT GCACGTTGCT GCAGCTGATA AA - #GACAGTTA        6360                                                                           - TCTGTTAATG GAGATGCTAC TAGATAGCGG GGCAGATCCA AATATAAAAT GC - #GCAAACGG        6420                                                                           - TTTTACTCCT TTGTTTAATG CAGTATATGA TCATAACCGT ATAAAGTTAT TA - #TTTCTTTA        6480                                                                           - CGGGGCTGAT ATCAATATTA CTGACTCTTA CGGAAATACT CCTCTTACTT AT - #ATGACTAA        6540                                                                           - TTTTGATAAT AAATATGTAA ATTCAATAAT TATCTTACAA ATATATCTAC TT - #AAAAAAGA        6600                                                                           - ATATAACGAT GAAAGATTGT TTCCACCTGG TATGATAAAA AATTTAAACT TT - #ATAGAATC        6660                                                                           - AAACGATAGT CTTAAAGTTA TAGCTAAAAA GTGTAATTCG TTAATACGCT AT - #AAGAAAAA        6720                                                                           - TAAAGACATA GATGCAGATA ACGTATTATT GGAGCTTTTA GAGGAAGAGG AA - #GAAGATGA        6780                                                                           - AATAGACAGA TGGCATACTA CATGTAAAAT ATCTTAAATA GTAATTAAAT CA - #TTGAAATA        6840                                                                           - TTAACTTACA AGATGATCGA GGTCACTTAT TATACTCTTT AATAATGGGT AC - #AAAGAGTA        6900                                                                           - TTCATACGTT AGTTAAATCT AACGATGTAA TACGTGTTCG TGAATTAATA AA - #GGATGATA        6960                                                                           - GATGTTTGAT AAATAAAAGA AATAGAAGAA ATCAGTCACC TGTATATATA GC - #TATATACA        7020                                                                           - AAGGACTTTA TGAAATGACT GAAATGTTAT TGCTAAATAA TGCAAGTCTA GA - #TACTAAAA        7080                                                                           - TACCTTCTTT AATTATAGCA GCTAAAAATA ATGACTTACC TATGATAAAA TT - #ATTGATAC        7140                                                                           - AATACGGGGC AAAATTAAAT GATATTTATT TAAGGGACAC AGCATTAATG AT - #AGCTCTCA        7200                                                                           - GAAATGGTTA CCTAGATATA GCTGAATATT TACTTTCATT AGGAGCAGAA TT - #TGTTAAAT        7260                                                                           - ACAGACATAA GGTAATATAT AAATATCTAT CAAAAGATGC GTATGAATTA CT - #TTTTAGAT        7320                                                                           #        7351      TATA ATAGATTGAG A                                           - (2) INFORMATION FOR SEQ ID NO:40:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 7091 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:                                 - AGATATTTGT TAGCTTCTGC CGGAGATACC GTGAAAATCT ATTTTCTGGA AG - #GAAAGGGA          60                                                                           - GGTCTTATCT ATTCTGTCAG CAGAGTAGGT TCCTCTAATG ACGAAGACAA TA - #GTGAATAC         120                                                                           - TTGCATGAAG GTCACTGTGT AGAGTTCAAA ACTGATCATC AGTGTTTGAT AA - #CTCTAGCG         180                                                                           - TGTACGAGTC CTTCTAACAC TGTGGTTTAT TGGCTGGAAT AAAAGGATAA AG - #ACACCTAT         240                                                                           - ACTGATTCAT TTTCATCTGT CAACGTTTCT CTAAGAGATT CATAGGTATT AT - #TATTACAT         300                                                                           - CGATCTAGAA GTCTAATAAC TGCTAAGTAT ATTATTGGAT TTAACGCGCT AT - #AAACGCAT         360                                                                           - CCAAAACCTA CAAATATAGG AGAAGCTTCT CTTATGAAAC TTCTTAAAGC TT - #TACTCTTA         420                                                                           - CTATTACTAC TCAAAAGAGA TATTACATTA ATTATGTGAT GAGGCATCCA AC - #ATATAAAG         480                                                                           - AAGACTAAAG CTGTAGAAGC TGTTATGAAG AATATCTTAT CAGATATATT AG - #ATGCATTG         540                                                                           - TTAGTTCTGT AGATCAGTAA CGTATAGCAT ACGAGTATAA TTATCGTAGG TA - #GTAGGTAT         600                                                                           - CCTAAAATAA ATCTGATACA GATAATAACT TTGTAAATCA ATTCAGCAAT TT - #CTCTATTA         660                                                                           - TCATGATAAT GATTAATACA CAGCGTGTCG TTATTTTTTG TTACGATAGT AT - #TTCTAAAG         720                                                                           - TAAAGAGCAG GAATCCCTAG TATAATAGAA ATAATCCATA TGAAAAATAT AG - #TAATGTAC         780                                                                           - ATATTTCTAA TGTTAACATA TTTATAGGTA AATCCAGGAA GGGTAATTTT TA - #CATATCTA         840                                                                           - TATACGCTTA TTACAGTTAT TAAAAATATA CTTGCAAACA TGTTAGAAGT AA - #AAAAGAAA         900                                                                           - GAACTAATTT TACAAAGTGC TTTACCAAAA TGCCAATGGA AATTACTTAG TA - #TGTATATA         960                                                                           - ATGTATAAAG GTATGAATAT CACAAACAGC AAATCGGCTA TTCCCAAGTT GA - #GAAACGGT        1020                                                                           - ATAATAGATA TATTTCTAGA TACCATTAAT AACCTTATAA GCTTGACGTT TC - #CTATAATG        1080                                                                           - CCTACTAAGA AAACTAGAAG ATACATACAT ACTAACGCCA TACGAGAGTA AC - #TACTCATC        1140                                                                           - GTATAACTAC TGTTGCTAAC AGTGACACTG ATGTTATAAC TCATCTTTGA TG - #TGGTATAA        1200                                                                           - ATGTATAATA ACTATATTAC ACTGGTATTT TATTTCAGTT ATATACTATA TA - #GTATTAAA        1260                                                                           - AATTATATTT GTATAATTAT ATTATTATAT TCAGTGTAGA AAGTAAAATA CT - #ATAAATAT        1320                                                                           - GTATCTCTTA TTTATAACTT ATTAGTAAAG TATGTACTAT TCAGTTATAT TG - #TTTTATAA        1380                                                                           - AAGCTAAATG CTACTAGATT GATATAAATG AATATGTAAT AAATTAGTAA TG - #TAGTATAC        1440                                                                           - TAATATTAAC TCACATTTGA CTAATTAGCT ATAAAAACCC GGGCTGCAGG AA - #TTCCTCGA        1500                                                                           - GACGCGTGGC ATGCAAGCTT ATAAAAATCA CAAGTCTCTG TCACTTTTTT TG - #TCTAGTTT        1560                                                                           - TTTTTTCTCC TCTTGGTTCA GACGTTCTCT TCTTCGTCGG AGTCTTTCAA GT - #GTCGGTAG        1620                                                                           - CCGTTTTTGC GGTGTCGCAG TCGGTCTAGC AGGTTGGGCT TCTGTCCCTT GT - #CCTGCGTG        1680                                                                           - CCAGTCTGTC CGTCCAAAGA ATCTGTACCG TTCTCGTGCG CTCGCTGCTC TG - #CGTCCAGA        1740                                                                           - CGGACCAGGG CCAGAAGCAT CTGGTAAGCC TGCTCGTTGG TGTAAGGCGG AG - #CCGCCGTG        1800                                                                           - GATGCATCAG ACGACGGTGG TCCCGGTCCT TTGCGACCAG AATTATAAAC AC - #TTTCCTCG        1860                                                                           - TAGGAAGGCG GAGCCTGTAA CGACGTGTCT TTGGTGTTGC CCGACGTCAC GG - #TGGTCCCG        1920                                                                           - TCGGCGGACA CCAGATAGGG AAAGAGGTTC TGCAGCGGCT GCATGCAGAG AC - #GCCGCTGT        1980                                                                           - CGAGTATAGA TCAAATAAAT GATAATGACG ACGGCTATGG CCACGAGGAT GA - #TGGTGAAG        2040                                                                           - GCTCCGAAGG GGTTTTTGAG GAAGGTGGCA ACGCCTTCGA CCACGGAGGC CA - #CCGCGCCA        2100                                                                           - CCCACGGCCC CAATGGCTAC GCCAACGGCC TTTCCCGCGG CGCCCAGGCC GC - #TCATGAGG        2160                                                                           - TCGTCCAGAC CCTTGAGGTA GGGCGGCAGC GGGTCGACTA CCTTGTCCTC CA - #CGTACTTT        2220                                                                           - ACCCGCTGCT TATACGAATT GAACTCGCGC ATGATCTCCT CGAGATCAAA AA - #CGTTGCTG        2280                                                                           - GAACGCAATT CTTTCTGCGA GTAAAGTTCC AGTACCCTGA AGTCGGTGTT TT - #CCAGCGGG        2340                                                                           - TCGATGTCTA GGGCGATCAT GCTGTCGACG GTGGAGATGC TGCTGAGGTC AA - #TCATGCGT        2400                                                                           - TTGAAGAGGT AGTCCACGTA CTCGTAGGCC GAGTTGCCGG CGATGAAGAT CT - #TGAGGCTG        2460                                                                           - GGAAGCTGAC ATTCCTCAGT GCGGTGGTTG CCCAACAGGA TTTCGTTATC CT - #CGCCCAGT        2520                                                                           - TGACCGTACT GCACGTACGA GCTGTTGGCG AAATTAAAGA TGACCACTGG TC - #GTGAGTAG        2580                                                                           - CAGCGTCCTG GCGATTCCTT CACATTCATA TCACGCAGCA CCTTGACGCT GG - #TTTGGTTA        2640                                                                           - ATGGTCACGC AGCTGGCCAG ACCCAGGACA TCACCCATGA AACGCGCGGC AA - #TCGGTTTG        2700                                                                           - TTGTAGATGG CCGAGAGAAT AGCTGACGGG TTGATCTTGC TAAGTTCCTT GA - #AGACCTCT        2760                                                                           - AGGGTGCGCC GTTGATCCAC ACACCAGGCT TCTGCGATTT CGGCCAGCGC CC - #GGTTGATG        2820                                                                           - TAACCGCGCA ACGTGTCATA GGTGAACTGC AGCTGGGCGT AGACCAGATT GT - #GCACCGAC        2880                                                                           - TCCATGTTGG ATAAATGAGT TGCATTGTTG CCATCTGTAC TTCTTTTGGT TC - #TATTATGA        2940                                                                           - GTAAGATTCA GACTGGAGCG GTTGGCCAAA CGTTCGAGTT CCACCAGAGA TT - #TTTGCTTG        3000                                                                           - ATACCTTGCC AGAACACCAC CAAACCACCA GTGGTTTCAA AGACGGACAC GT - #TTCCATAT        3060                                                                           - TTTTCATATG TTTGATTGTA TGAAGTATTG AAAATCTGCT GTAACTTATT TA - #TGGCCTCA        3120                                                                           - TCACGTACAC AGTCCAGCGC AGAGTCGGAC ATGTTCACCT CTTGCTTCTT AG - #ATAAGAAA        3180                                                                           - GTGGCGGTCA TTTTGGCAGA AGAAAAGTGA TACGAGTCCT CGGCTTCGGA AC - #GAATGGTG        3240                                                                           - CGTTCCGAGG CTTCCCAGAA AGTGAGTTGA CAAGTAACAT TCTTCTCGTC CT - #GTATATCC        3300                                                                           - CAGGAGATCA CTGAGTCCGC ACGTTCAAGA AAAGCCACCA ACCTGTGGGT CT - #CTAACGCA        3360                                                                           - GAATTCGGTC TTTCAAAGTC GGAGACGATA GTGTAGTTCG GAAAAATGAA AA - #ACTTGTCG        3420                                                                           - GCGTTTTCTC CAAAATAGCT GGCATTGCGA TTAGTTCCGT TGTAGAAAGG AG - #AAATGTCA        3480                                                                           - ACCACATCAC CCGTGGAAGT TGCGAAAAAA TGATAGGGAT ACTTGGAGCG CG - #CAGTAGTG        3540                                                                           - ATGGTCACCA TACAATTCAG ATTACAGGTC TCACGATAGA GCCAGGTGCT GC - #CGCGGCTG        3600                                                                           - TGCCATTGAT CCTTGACCGT CACGTAACGG GTACTGTGGG TGTTGGAATA AT - #CGTCGGGC        3660                                                                           - ATTAATTGCA TGGTTTTGTT TTCATAGCTG TCCCTATGAT AAGCCACGAA AA - #CCGTGCCT        3720                                                                           - GCTATAACGC GGCTGTAGGA ACTGTAGCAC TGACTGTGAC TGTTGATATG AT - #GAATCTCC        3780                                                                           - CACATAGGAG GCGCCACGTA TTCCGTGTTG CTGCCCAGCA GATAAGTGGT GT - #GGATGTAA        3840                                                                           - GCGTAGCTAC GACGAAACGT CAAAACCTTC TGGTAGACTC GTACCTTAAA GG - #TGTGCGCG        3900                                                                           - ACGATGTTGC GTTTGTAGAC CACCATGATG CCCTCGTCCA GGTCTTCATT GA - #TGGGCTTC        3960                                                                           - ATCGAGGTGC AGACGATATT ACGTTCAAAG CGAATAAGAT CCGTACCCTG AG - #CCATAGAA        4020                                                                           - CACACGCGAT AGGGGTACTT GGTGGTGTTG ACCCCCACCA CATCTCCGTA CT - #TGAGGGTA        4080                                                                           - GTGTTGTAGA TGGTCTCGTT AACACCATGG CTGACCGTTT GGGAAGAAGT TA - #CGCGTTGA        4140                                                                           - GAGACTGAAC CGGATCGAGA ATGAGCAGCA GACGTCGTAT GAGAGGAATG GT - #GACTGTGA        4200                                                                           - GTAGCAGAAG TTCCACGAGT AGAAGATGAG GAAACCGCAG CACCCAGACA GA - #CGATACAC        4260                                                                           - AAGTTAACGC AGACTACCAG GCACCAGATC CTGGATTCCA TTACGATACA AA - #CTTAACGG        4320                                                                           - ATATCGCGAT AATGAAATAA TTTATGATTA TTTCTCGCTT TCAATTTAAC AC - #AACCCTCA        4380                                                                           - AGAACCTTTG TATTTATTTT CACTTTTTAA GTATAGAATA AAGAAGCTCT AA - #TTAATTAA        4440                                                                           - GCTACAAATA GTTTCGTTTT CACCTTGTCT AATAACTAAT TAATTAACCC GG - #ATCCCGAT        4500                                                                           - TTTTATGACT AGTTAATCAA ATAAAAAGCA TACAAGCTAT TGCTTCGCTA TC - #GTTACAAA        4560                                                                           - ATGGCAGGAA TTTTGTGTAA ACTAAGCCAC ATACTTGCCA ATGAAAAAAA TA - #GTAGAAAG        4620                                                                           - GATACTATTT TAATGGGATT AGATGTTAAG GTTCCTTGGG ATTATAGTAA CT - #GGGCATCT        4680                                                                           - GTTAACTTTT ACGACGTTAG GTTAGATACT GATGTTACAG ATTATAATAA TG - #TTACAATA        4740                                                                           - AAATACATGA CAGGATGTGA TATTTTTCCT CATATAACTC TTGGAATAGC AA - #ATATGGAT        4800                                                                           - CAATGTGATA GATTTGAAAA TTTCAAAAAG CAAATAACTG ATCAAGATTT AC - #AGACTATT        4860                                                                           - TCTATAGTCT GTAAAGAAGA GATGTGTTTT CCTCAGAGTA ACGCCTCTAA AC - #AGTTGGGA        4920                                                                           - GCGAAAGGAT GCGCTGTAGT TATGAAACTG GAGGTATCTG ATGAACTTAG AG - #CCCTAAGA        4980                                                                           - AATGTTCTGC TGAATGCGGT ACCCTGTTCG AAGGACGTGT TTGGTGATAT CA - #CAGTAGAT        5040                                                                           - AATCCGTGGA ATCCTCACAT AACAGTAGGA TATGTTAAGG AGGACGATGT CG - #AAAACAAG        5100                                                                           - AAACGCCTAA TGGAGTGCAT GTCCAAGTTT AGGGGGCAAG AAATACAAGT TC - #TAGGATGG        5160                                                                           - TATTAATAAG TATCTAAGTA TTTGGTATAA TTTATTAAAT AGTATAATTA TA - #ACAAATAA        5220                                                                           - TAAATAACAT GATAACGGTT TTTATTAGAA TAAAATAGAG ATAATATCAT AA - #TGATATAT        5280                                                                           - AATACTTCAT TACCAGAAAT GAGTAATGGA AGACTTATAA ATGAACTGCA TA - #AAGCTATA        5340                                                                           - AGGTATAGAG ATATAAATTT AGTAAGGTAT ATACTTAAAA AATGCAAATA CA - #ATAACGTA        5400                                                                           - AATATACTAT CAACGTCTTT GTATTTAGCC GTAAGTATTT CTGATATAGA AA - #TGGTAAAA        5460                                                                           - TTATTACTAG AACACGGTGC CGATATTTTA AAATGTAAAA ATCCTCCTCT TC - #ATAAAGCT        5520                                                                           - GCTAGTTTAG ATAATACAGA AATTGCTAAA CTACTAATAG ATTCTGGCGC TG - #ACATAGAA        5580                                                                           - CAGATACATT CTGGAAATAG TCCGTTATAT ATTTCTGTAT ATAGAAACAA TA - #AGTCATTA        5640                                                                           - ACTAGATATT TATTAAAAAA AGGTGTTAAT TGTAATAGAT TCTTTCTAAA TT - #ATTACGAT        5700                                                                           - GTACTGTATG ATAAGATATC TGATGATATG TATAAAATAT TTATAGATTT TA - #ATATTGAT        5760                                                                           - CTTAATATAC AAACTAGAAA TTTTGAAACT CCGTTACATT ACGCTATAAA GT - #ATAAGAAT        5820                                                                           - ATAGATTTAA TTAGGATATT GTTAGATAAT AGTATTAAAA TAGATAAAAG TT - #TATTTTTG        5880                                                                           - CATAAACAGT ATCTCATAAA GGCACTTAAA AATAATTGTA GTTACGATAT AA - #TAGCGTTA        5940                                                                           - CTTATAAATC ACGGAGTGCC TATAAACGAA CAAGATGATT TAGGTAAAAC CC - #CATTACAT        6000                                                                           - CATTCGGTAA TTAATAGAAG AAAAGATGTA ACAGCACTTC TGTTAAATCT AG - #GAGCTGAT        6060                                                                           - ATAAACGTAA TAGATGACTG TATGGGCAGT CCCTTACATT ACGCTGTTTC AC - #GTAACGAT        6120                                                                           - ATCGAAACAA CAAAGACACT TTTAGAAAGA GGATCTAATG TTAATGTGGT TA - #ATAATCAT        6180                                                                           - ATAGATACCG TTCTAAATAT AGCTGTTGCA TCTAAAAACA AAACTATAGT AA - #ACTTATTA        6240                                                                           - CTGAAGTACG GTACTGATAC AAAGTTGGTA GGATTAGATA AACATGTTAT TC - #ACATAGCT        6300                                                                           - ATAGAAATGA AAGATATTAA TATACTGAAT GCGATCTTAT TATATGGTTG CT - #ATGTAAAC        6360                                                                           - GTCTATAATC ATAAAGGTTT CACTCCTCTA TACATGGCAG TTAGTTCTAT GA - #AAACAGAA        6420                                                                           - TTTGTTAAAC TCTTACTTGA CCACGGTGCT TACGTAAATG CTAAAGCTAA GT - #TATCTGGA        6480                                                                           - AATACTCCTT TACATAAAGC TATGTTATCT AATAGTTTTA ATAATATAAA AT - #TACTTTTA        6540                                                                           - TCTTATAACG CCGACTATAA TTCTCTAAAT AATCACGGTA ATACGCCTCT AA - #CTTGTGTT        6600                                                                           - AGCTTTTTAG ATGACAAGAT AGCTATTATG ATAATATCTA AAATGATGTT AG - #AAATATCT        6660                                                                           - AAAAATCCTG AAATAGCTAA TTCAGAAGGT TTTATAGTAA ACATGGAACA TA - #TAAACAGT        6720                                                                           - AATAAAAGAC TACTATCTAT AAAAGAATCA TGCGAAAAAG AACTAGATGT TA - #TAACACAT        6780                                                                           - ATAAAGTTAA ATTCTATATA TTCTTTTAAT ATCTTTCTTG ACAATAACAT AG - #ATCTTATG        6840                                                                           - GTAAAGTTCG TAACTAATCC TAGAGTTAAT AAGATACCTG CATGTATACG TA - #TATATAGG        6900                                                                           - GAATTAATAC GGAAAAATAA ATCATTAGCT TTTCATAGAC ATCAGCTAAT AG - #TTAAAGCT        6960                                                                           - GTAAAAGAGA GTAAGAATCT AGGAATAATA GGTAGGTTAC CTATAGATAT CA - #AACATATA        7020                                                                           - ATAATGGAAC TATTAAGTAA TAATGATTTA CATTCTGTTA TCACCAGCTG TT - #GTAACCCA        7080                                                                           #     7091                                                                     - (2) INFORMATION FOR SEQ ID NO:41:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 4768 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:                                 - AAGCTTGCGG CCGCTCATTA GACAAGCGAA TGAGGGACGA AAACGTGGAG GA - #GGTATTAA          60                                                                           - GTTTGGAGAA ATGGAGAGAG ACTGTTTAAT AGCGCATGGC GCAGCCAATA CT - #ATTACAGA         120                                                                           - AGTTTTGAAA GATTCGGAAG AAGATTATCA AGATGTGTAT GTTTGTGAAA AT - #TGTGGAGA         180                                                                           - CATAGCAGCA CAAATCAAGG GTATTAATAC ATGTCTTAGA TGTTCAAAAC TT - #AATCTCTC         240                                                                           - TCCTCTCTTA ACAAAAATTG ATACCACGCA CGTATCTAAA GTATTTCTTA CT - #CAAATGAA         300                                                                           - CGCCAGAGGC GTAAAAGTCA AATTAGATTT CGAACGAAGG CCTCCTTCGT TT - #TATAAACC         360                                                                           - ATTAGATAAA GTTGATCTCA AGCCGTCTTT TCTGGTGTAA TAAAAATTAA TT - #AATTACTC         420                                                                           - GAGGGTACCG GATCCCCCAG CTTATAAAAA TCACAAGTCT CTGACACTTT TT - #TTGTCTAG         480                                                                           - TTTTTTTTTC TCCTCTTGGT TCAGACGTTC TCTTCTTCGT CGGAGTCTTT CA - #AGTGTCGG         540                                                                           - TAGCCGTTTT TGCGGTGTCG CAGTCGGTCT AGCAGGTTGG GCTTCTGTCC CT - #TGTCCTGC         600                                                                           - GTGCCAGTCT GTCCGTCCAA AGAATCTGTA CCGTTCTCGT GCGCTCGCTG CT - #CTGCGTCC         660                                                                           - AGACGGACCA GGGCCAGAAG CATCTGGTAA GCCTGCTCGT TGGTGTAAGG CG - #GAGCCGCC         720                                                                           - GTGGATGCAT CAGACGACGG TGGTCCCGGT CCTTTGCGAC CAGAATTATA AA - #CACTTTCC         780                                                                           - TCGTAGGAAG GCGGAGCCTG TAACGACGTG TCTTTGGTGT TGCCCGACGT CA - #CGGTGGTC         840                                                                           - CCGTCGGCGG ACACCAGATA GGGAAAGAGG TTCTGCAGCG GCTGCATGCA GA - #GACGCCGC         900                                                                           - TGTCGAGTAT AGATCAAATA AATGATAATG ACGACGGCTA TGGCCACGAG GA - #TGATGGTG         960                                                                           - AAGGCTCCGA AGGGGTTTTT GAGGAAGGTG GCAACGCCTT CGACCACGGA GG - #CCACCGCG        1020                                                                           - CCACCCACGG CCCCAATGGC TACGCCAACG GCCTTTCCCG CGGCGCCCAG GC - #CGCTCATG        1080                                                                           - AGGTCGTCCA GACCCTTGAG GTAGGGCGGC AGCGGGTCGA CTACCTTGTC CT - #CCACGTAC        1140                                                                           - TTTACCCGCT GCTTATACGA ATTGAACTCG CGCATGATCT CCTCGAGATC AA - #AAACGTTG        1200                                                                           - CTGGAACGCA ATTCTTTCTG CGAGTAAAGT TCCAGTACCC TGAAGTCGGT GT - #TTTCCAGC        1260                                                                           - GGGTCGATGT CTAGGGCGAT CATGCTGTCG ACGGTGGAGA TGCTGCTGAG GT - #CAATCATG        1320                                                                           - CGTTTGAAGA GGTAGTCCAC GTACTCGTAG GCCGAGTTGC CGGCGATGAA GA - #TCTTGAGG        1380                                                                           - CTGGGAAGCT GACATTCCTC AGTGCGGTGG TTGCCCAACA GGATTTCGTT AT - #CCTCGCCC        1440                                                                           - AGTTGACCGT ACTGCACGTA CGAGCTGTTG GCGAAATTAA AGATGACCAC TG - #GTCGTGAG        1500                                                                           - TAGCAGCGTC CTGGCGATTC CTTCACATTC ATATCACGCA GCACCTTGAC GC - #TGGTTTGG        1560                                                                           - TTAATGGTCA CGCAGCTGGC CAGACCCAGG ACATCACCCA TGAAACGCGC GG - #CAATCGGT        1620                                                                           - TTGTTGTAGA TGGCCGAGAG AATAGCTGAC GGGTTGATCT TGCTAAGTTC CT - #TGAAGACC        1680                                                                           - TCTAGGGTGC GCCGTTGATC CACACACCAG GCTTCTGCGA TTTCGGCCAG CG - #CCCGGTTG        1740                                                                           - ATGTAACCGC GCAACGTGTC ATAGGTGAAC TGCAGCTGGG CGTAGACCAG AT - #TGTGCACC        1800                                                                           - GACTCCATGT TGGATAAATG AGTTGCATTG TTGCCATCTG TACTTCTTTT GG - #TTCTATTA        1860                                                                           - TGAGTAAGAT TCAGACTGGA GCGGTTGGCC AAACGTTCGA GTTCCACCAG AG - #ATTTTTGC        1920                                                                           - TTGATACCTT GCCAGAACAC CACCAAACCA CCAGTGGTTT CAAAGACGGA CA - #CGTTTCCA        1980                                                                           - TATTTTTCAT ATGTTTGATT GTATGAAGTA TTGAAAATCT GCTGTAACTT AT - #TTATGGCC        2040                                                                           - TCATCACGTA CACAGTCCAG CGCAGAGTCG GACATGTTCA CCTCTTGCTT CT - #TAGATAAG        2100                                                                           - AAAGTGGCGG TCATTTTGGC AGAAGAAAAG TGATACGAGT CCTCGGCTTC GG - #AACGAATG        2160                                                                           - GTGCGTTCCG AGGCTTCCCA GAAAGTGAGT TGACAAGTAA CATTCTTCTC GT - #CCTGTATA        2220                                                                           - TCCCAGGAGA TCACTGAGTC CGCACGTTCA AGAAAAGCCA CCAACCTGTG GG - #TCTCTAAC        2280                                                                           - GCAGAATTCG GTCTTTCAAA GTCGGAGACG ATAGTGTAGT TCGGAAAAAT GA - #AAAACTTG        2340                                                                           - TCGGCGTTTT CTCCAAAATA GCTGGCATTG CGATTAGTTC CGTTGTAGAA AG - #GAGAAATG        2400                                                                           - TCAACCACAT CACCCGTGGA AGTTGCGAAA AAATGATAGG GATACTTGGA GC - #GCGCAGTA        2460                                                                           - GTGATGGTCA CCATACAATT CAGATTACAG GTCTCACGAT AGAGCCAGGT GC - #TGCCGCGG        2520                                                                           - CTGTGCCATT GATCCTTGAC CGTCACGTAA CGGGTACTGT GGGTGTTGGA AT - #AATCGTCG        2580                                                                           - GGCATTAATT GCATGGTTTT GTTTTCATAG CTGTCCCTAT GATAAGCCAC GA - #AAACCGTG        2640                                                                           - CCTGCTATAA CGCGGCTGTA GGAACTGTAG CACTGACTGT GACTGTTGAT AT - #GATGAATC        2700                                                                           - TCCCACATAG GAGGCGCCAC GTATTCCGTG TTGCTGCCCA GCAGATAAGT GG - #TGTGGATG        2760                                                                           - TAAGCGTAGC TACGACGAAA CGTCAAAACC TTCTGGTAGA CTCGTACCTT AA - #AGGTGTGC        2820                                                                           - GCGACGATGT TGCGTTTGTA GACCACCATG ATGCCCTCGT CCAGGTCTTC AT - #TGATGGGC        2880                                                                           - TTCATCGAGG TGCAGACGAT ATTACGTTCA AAGCGAATAA GATCCGTACC CT - #GAGCCATA        2940                                                                           - GAACACACGC GATAGGGGTA CTTGGTGGTG TTGACCCCCA CCACATCTCC GT - #ACTTGAGG        3000                                                                           - GTAGTGTTGT AGATGGTCTC GTTAACACCA TGGCTGACCG TTTGGGAAGA AG - #TTACGCGT        3060                                                                           - TGAGAGACTG AACCGGATCG AGAATGAGCA GCAGACGTCG TATGAGAGGA AT - #GGTGACTG        3120                                                                           - TGAGTAGCAG AAGTTCCACG AGTAGAAGAT GAGGAAACCG CAGCACCCAG AC - #AGACGATA        3180                                                                           - CACAAGTTAA CGCAGACTAC CAGGCACCAG ATCCTGGATT CCATTACGAT AC - #AAACTTAA        3240                                                                           - CGGATATCGC GATAATGAAA TAATTTATGA TTATTTCTCG CTTTCAATTT AA - #CACAACCC        3300                                                                           - TCAAGAACCT TTGTATTTAT TTTCACTTTT TAAGTATAGA ATAAAGAAGC TG - #GGAATCGA        3360                                                                           - TTCGCGATAG CTGATTAGTT TTTGTTAACA AAAATGTGGG AGAATCTAAT TA - #GTTTTTCT        3420                                                                           - TTACACAATT GACGTACATG AGTCTGAGTT CCTTGTTTTT GCTAATTATT TC - #ATCCAATT        3480                                                                           - TATTATTCTT GACGATATCG AGATCTTTTG TATAGGAGTC AGACTTGTAT TC - #AACATGCT        3540                                                                           - TTTCTATAAT CATCTTAGTT ATTTCGGCAT CATCCAATAG TACATTTTCC AG - #ATTAACAG        3600                                                                           - AGTAGATATT AATGTCGTAT TTGAACAGAG CCTGTAACAT CTCAATGTCT TT - #ATTATCTA        3660                                                                           - TAGCCAATTT AATGTCCGGA ATGAAGAGAA GGGAATTATT GGTGTTTGTC GA - #CGTCATAT        3720                                                                           - AGTCGAGCAA GAGAATCATC ATATCCACGT GTCCATTTTT TATAGTGGTG TG - #AATACAAC        3780                                                                           - TAAGGAGAAT AGCCAGATCA AAAGTAGATG GTATTTCTGA AAGAAAGTAT GA - #TACAATAC        3840                                                                           - TTACATCATT AAGCATGACG GCATGATAAA ATGAAGTTTT CCATCCAGTT TT - #CCCATAGA        3900                                                                           - ACATCAGTCT CCAATTTTTC TTAAACAGTT TCACCGTTTG CATGTTACCA CT - #ATCAACCG        3960                                                                           - CATAATACAA TGCGGTGTTT CCTTTGTCAT CAAATTGTGA ATCATCCATT CC - #ACTGAATA        4020                                                                           - GCAAAATCTT TACTATTTTG GTATCTTCTA ATGTGGCTGC CTGATGTAAT GG - #AAATTCAT        4080                                                                           - TCTCTAGAAG ATTTTTCAAT GCTCCAGCGT TCAACAACGT ACATACTAGA CG - #CACGTTAT        4140                                                                           - TATCAGCTAT TGCATAATAC AAGGCACTAT GTCCATGGAC ATCCGCCTTA AA - #TGTATCTT        4200                                                                           - TACTAGAGAG AAAGCTTTTC AGCTGCTTAG ACTTCCAAGT ATTAATTCGT GA - #CAGATCCA        4260                                                                           - TGTCTGAAAC GAGACGCTAA TTAGTGTATA TTTTTTCATT TTTTATAATT TT - #GTCATATT        4320                                                                           - GCACCAGAAT TAATAATATC TCTAATAGAT CTAATTTAAT TTAATTTATA TA - #ACTTATTT        4380                                                                           - TTTGAATATA CTTTTAATTA ACAAAAGAGT TAAGTTACTC ATATGGACGC CG - #TCCAGTCT        4440                                                                           - GAACATCAAT CTTTTTAGCC AGAGATATCA TAGCCGCTCT TAGAGTTTCA GC - #GTGATTTT        4500                                                                           - CCAACCTAAA TAGAACTTCA TCGTTGCGTT TACAACACTT TTCTATTTGT TC - #AAACTTTG        4560                                                                           - TTGTTACATT AGTAATCTTT TTTTCCAAAT TAGTTAGCCG TTGTTTGAGA GT - #TTCCTCAT        4620                                                                           - TGTCGTCTTC ATCGGCTTTA ACAATTGCTT CGCGTTTAGC CTCCTGGCTG TT - #CTTATCAG        4680                                                                           - CCTTTGTAGA AAAAAATTCA GTTGCTGGAA TTGCAAGATC GTCATCTCCG GG - #GAAAAGAG        4740                                                                           #           4768   CCGC GGGAATTC                                               - (2) INFORMATION FOR SEQ ID NO:42:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 2550 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:                                 - ATGGAATCCA GGATCTGGTG CCTGGTAGTC TGCGTTAACT TGTGTATCGT CT - #GTCTGGGT          60                                                                           - GCTGCGGTTT CCTCATCTTC TACTCGTGGA ACTTCTGCTA CTCACAGTCA CC - #ATTCCTCT         120                                                                           - CATACGACGT CTGCTGCTCA TTCTCGATCC GGTTCAGTCT CTCAACGCGT AA - #CTTCTTCC         180                                                                           - CAAACGGTCA GCCATGGTGT TAACGAGACC ATCTACAACA CTACCCTCAA GT - #ACGGAGAT         240                                                                           - GTGGTGGGGG TCAACACCAC CAAGTACCCC TATCGCGTGT GTTCTATGGC TC - #AGGGTACG         300                                                                           - GATCTTATTC GCTTTGAACG TAATATCGTC TGCACCTCGA TGAAGCCCAT CA - #ATGAAGAC         360                                                                           - CTGGACGAGG GCATCATGGT GGTCTACAAA CGCAACATCG TCGCGCACAC CT - #TTAAGGTA         420                                                                           - CGAGTCTACC AGAAGGTTTT GACGTTTCGT CGTAGCTACG CTTACATCCA CA - #CCACTTAT         480                                                                           - CTGCTGGGCA GCAACACGGA ATACGTGGCG CCTCCTATGT GGGAGATTCA TC - #ATATCAAC         540                                                                           - AGTCACAGTC AGTGCTACAG TTCCTACAGC CGCGTTATAG CAGGCACGGT TT - #TCGTGGCT         600                                                                           - TATCATAGGG ACAGCTATGA AAACAAAACC ATGCAATTAA TGCCCGACGA TT - #ATTCCAAC         660                                                                           - ACCCACAGTA CCCGTTACGT GACGGTCAAG GATCAATGGC ACAGCCGCGG CA - #GCACCTGG         720                                                                           - CTCTATCGTG AGACCTGTAA TCTGAATTGT ATGGTGACCA TCACTACTGC GC - #GCTCCAAG         780                                                                           - TATCCCTATC ATTTTTTCGC AACTTCCACG GGTGATGTGG TTGACATTTC TC - #CTTTCTAC         840                                                                           - AACGGAACTA ATCGCAATGC CAGCTATTTT GGAGAAAACG CCGACAAGTT TT - #TCATTTTT         900                                                                           - CCGAACTACA CTATCGTCTC CGACTTTGAA AGACCGAATT CTGCGTTAGA GA - #CCCACAGG         960                                                                           - TTGGTGGCTT TTCTTGAACG TGCGGACTCA GTGATCTCCT GGGATATACA GG - #ACGAGAAG        1020                                                                           - AATGTTACTT GTCAACTCAC TTTCTGGGAA GCCTCGGAAC GCACCATTCG TT - #CCGAAGCC        1080                                                                           - GAGGACTCGT ATCACTTTTC TTCTGCCAAA ATGACCGCCA CTTTCTTATC TA - #AGAAGCAA        1140                                                                           - GAGGTGAACA TGTCCGACTC TGCGCTGGAC TGTGTACGTG ATGAGGCCAT AA - #ATAAGTTA        1200                                                                           - CAGCAGATTT TCAATACTTC ATACAATCAA ACATATGAAA AATATGGAAA CG - #TGTCCGTC        1260                                                                           - TTTGAAACCA CTGGTGGTTT GGTGGTGTTC TGGCAAGGTA TCAAGCAAAA AT - #CTCTGGTG        1320                                                                           - GAACTCGAAC GTTTGGCCAA CCGCTCCAGT CTGAATCTTA CTCATAATAG AA - #CCAAAAGA        1380                                                                           - AGTACAGATG GCAACAATGC AACTCATTTA TCCAACATGG AGTCGGTGCA CA - #ATCTGGTC        1440                                                                           - TACGCCCAGC TGCAGTTCAC CTATGACACG TTGCGCGGTT ACATCAACCG GG - #CGCTGGCC        1500                                                                           - GAAATCGCAG AAGCCTGGTG TGTGGATCAA CGGCGCACCC TAGAGGTCTT CA - #AGGAACTT        1560                                                                           - AGCAAGATCA ACCCGTCAGC TATTCTCTCG GCCATCTACA ACAAACCGAT TG - #CCGCGCGT        1620                                                                           - TTCATGGGTG ATGTCCTGGG TCTGGCCAGC TGCGTGACCA TTAACCAAAC CA - #GCGTCAAG        1680                                                                           - GTGCTGCGTG ATATGAATGT GAAGGAATCG CCAGGACGCT GCTACTCACG AC - #CAGTGGTC        1740                                                                           - ATCTTTAATT TCGCCAACAG CTCGTACGTG CAGTACGGTC AACTGGGCGA GG - #ATAACGAA        1800                                                                           - ATCCTGTTGG GCAACCACCG CACTGAGGAA TGTCAGCTTC CCAGCCTCAA GA - #TCTTCATC        1860                                                                           - GCCGGCAACT CGGCCTACGA GTACGTGGAC TACCTCTTCA AACGCATGAT TG - #ACCTCAGC        1920                                                                           - AGCATCTCCA CCGTCGACAG CATGATCGCC CTAGACATCG ACCCGCTGGA AA - #ACACCGAC        1980                                                                           - TTCAGGGTAC TGGAACTTTA CTCGCAGAAA GAATTGCGTT CCAGCAACGT TT - #TTGATCTC        2040                                                                           - GAGGAGATCA TGCGCGAGTT CAATTCGTAT AAGCAGCGGG TAAAGTACGT GG - #AGGACAAG        2100                                                                           - GTAGTCGACC CGCTGCCGCC CTACCTCAAG GGTCTGGACG ACACTCGACA GC - #GGCGTCTC        2160                                                                           - TGCATGCAGC CGCTGCAGAA CCTCTTTCCC TATCTGGTGT CCGCCGACGG GA - #CCACCGTG        2220                                                                           - ACGTCGGGCA ACACCAAAGA CACGTCGTTA CAGGCTCCGC CTTCCTACGA GG - #AAAGTGTT        2280                                                                           - TATAATTCTG GTCGCAAAGG ACCGGGACCA CCGTCGTCTG ATGCATCCAC GG - #CGGCTCCG        2340                                                                           - CCTTACACCA ACGAGCAGGC TTACCAGATG CTTCTGGCCC TGGTCCGTCT GG - #ACGCAGAG        2400                                                                           - CAGCGAGCGC ACGAGAACGG TACAGATTCT TTGGACGGAC AGACTGGCAC GC - #AGGACAAG        2460                                                                           - GGACAGAAGC CCAACCTGCT AGACCGACTG CGACACCGCA AAAACGGCTA CC - #GACACTTG        2520                                                                           #         2550     AAGA GAACGTCTGA                                             - (2) INFORMATION FOR SEQ ID NO:43:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 4594 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:43:                                 - AAGCTTGCGG CCGCTCATTA GACAAGCGAA TGAGGGACGA AAACGTGGAG GA - #GGTATTAA          60                                                                           - GTTTGGAGAA ATGGAGAGAG ACTGTTTAAT AGCGCATGGC GCAGCCAATA CT - #ATTACAGA         120                                                                           - AGTTTTGAAA GATTCGGAAG AAGATTATCA AGATGTGTAT GTTTGTGAAA AT - #TGTGGAGA         180                                                                           - CATAGCAGCA CAAATCAAGG GTATTAATAC ATGTCTTAGA TGTTCAAAAC TT - #AATCTCTC         240                                                                           - TCCTCTCTTA ACAAAAATTG ATACCACGCA CGTATCTAAA GTATTTCTTA CT - #CAAATGAA         300                                                                           - CGCCAGAGGC GTAAAAGTCA AATTAGATTT CGAACGAAGG CCTCCTTCGT TT - #TATAAACC         360                                                                           - ATTAGATAAA GTTGATCTCA AGCCGTCTTT TCTGGTGTAA TAAAAATTAA TT - #AATTACTC         420                                                                           - GAGGGTACCG GATCCCCCAG CTTATAAAAA TCACAAGACT CTGTCACTTT TT - #TTGACTAG         480                                                                           - TTTTTTTTTC TCCTCTTGGT TCAGACGTTC TCTTCTTCGT CGGAGTCTTT CA - #AGTGTCGG         540                                                                           - TAGCCGTTTT TGCGGTGTCG CAGTCGGTCT AGCAGGTTGG GCTTCTGTCC CT - #TGTCCTGC         600                                                                           - GTGCCAGTCT GTCCGTCCAA AGAATCTGTA CCGTTCTCGT GCGCTCGCTG CT - #CTGCGTCC         660                                                                           - AGACGGACCA GGGCCAGAAG CATCTGGTAA GCCTGCTCGT TGGTGTAAGG CG - #GAGCCGCC         720                                                                           - GTGGATGCAT CAGACGACGG TGGTCCCGGT CCTTTGCGAC CAGAATTATA AA - #CACTTTCC         780                                                                           - TCGTAGGAAG GCGGAGCCTG TAACGACGTG TCTTTGGTGT TGCCCGACGT CA - #CGGTGGTC         840                                                                           - CCGTCGGCGG ACACCAGATA GGGAAAGAGG TTCTGCAGCG GCTGCATGCA GA - #GACGCCGC         900                                                                           - TGTCGAGTGT CGTCCAGACC CTTGAGGTAG GGCGGCAGCG GGTCGACTAC CT - #TGTCCTCC         960                                                                           - ACGTACTTTA CCCGCTGCTT ATACGAATTG AACTCGCGCA TGATCTCCTC GA - #GATCAAAA        1020                                                                           - ACGTTGCTGG AACGCAATTC TTTCTGCGAG TAAAGTTCCA GTACCCTGAA GT - #CGGTGTTT        1080                                                                           - TCCAGCGGGT CGATGTCTAG GGCGATCATG CTGTCGACGG TGGAGATGCT GC - #TGAGGTCA        1140                                                                           - ATCATGCGTT TGAAGAGGTA GTCCACGTAC TCGTAGGCCG AGTTGCCGGC GA - #TGAAGATC        1200                                                                           - TTGAGGCTGG GAAGCTGACA TTCCTCAGTG CGGTGGTTGC CCAACAGGAT TT - #CGTTATCC        1260                                                                           - TCGCCCAGTT GACCGTACTG CACGTACGAG CTGTTGGCGA AATTAAAGAT GA - #CCACTGGT        1320                                                                           - CGTGAGTAGC AGCGTCCTGG CGATTCCTTC ACATTCATAT CACGCAGCAC CT - #TGACGCTG        1380                                                                           - GTTTGGTTAA TGGTCACGCA GCTGGCCAGA CCCAGGACAT CACCCATGAA AC - #GCGCGGCA        1440                                                                           - ATCGGTTTGT TGTAGATGGC CGAGAGAATA GCTGACGGGT TGATCTTGCT AA - #GTTCCTTG        1500                                                                           - AAGACCTCTA GGGTGCGCCG TTGATCCACA CACCAGGCTT CTGCGATTTC GG - #CCAGCGCC        1560                                                                           - CGGTTGATGT AACCGCGCAA CGTGTCATAG GTGAACTGCA GCTGGGCGTA GA - #CCAGATTG        1620                                                                           - TGCACCGACT CCATGTTGGA TAAATGAGTT GCATTGTTGC CATCTGTACT TC - #TTTTGGTT        1680                                                                           - CTATTATGAG TAAGATTCAG ACTGGAGCGG TTGGCCAAAC GTTCGAGTTC CA - #CCAGAGAT        1740                                                                           - TTTTGCTTGA TACCTTGCCA GAACACCACC AAACCACCAG TGGTTTCAAA GA - #CGGACACG        1800                                                                           - TTTCCATATT TTTCATATGT TTGATTGTAT GAAGTATTGA AAATCTGCTG TA - #ACTTATTT        1860                                                                           - ATGGCCTCAT CACGTACACA GTCCAGCGCA GAGTCGGACA TGTTCACCTC TT - #GCTTCTTA        1920                                                                           - GATAAGAAAG TGGCGGTCAT TTTGGCAGAA GAAAAGTGAT ACGAGTCCTC GG - #CTTCGGAA        1980                                                                           - CGAATGGTGC GTTCCGAGGC TTCCCAGAAA GTGAGTTGAC AAGTAACATT CT - #TCTCGTCC        2040                                                                           - TGTATATCCC AGGAGATCAC TGAGTCCGCA CGTTCAAGAA AAGCCACCAA CC - #TGTGGGTC        2100                                                                           - TCTAACGCAG AATTCGGTCT TTCAAAGTCG GAGACGATAG TGTAGTTCGG AA - #AAATGAAA        2160                                                                           - AACTTGTCGG CGTTTTCTCC AAAATAGCTG GCATTGCGAT TAGTTCCGTT GT - #AGAAAGGA        2220                                                                           - GAAATGTCAA CCACATCACC CGTGGAAGTT GCGAAAAAAT GATAGGGATA CT - #TGGAGCGC        2280                                                                           - GCAGTAGTGA TGGTCACCAT ACAATTCAGA TTACAGGTCT CACGATAGAG CC - #AGGTGCTG        2340                                                                           - CCGCGGCTGT GCCATTGATC CTTGACCGTC ACGTAACGGG TACTGTGGGT GT - #TGGAATAA        2400                                                                           - TCGTCGGGCA TTAATTGCAT GGTTTTGTTT TCATAGCTGT CCCTATGATA AG - #CCACGAAA        2460                                                                           - ACCGTGCCTG CTATAACGCG GCTGTAGGAA CTGTAGCACT GACTGTGACT GT - #TGATATGA        2520                                                                           - TGAATCTCCC ACATAGGAGG CGCCACGTAT TCCGTGTTGC TGCCCAGCAG AT - #AAGTGGTG        2580                                                                           - TGGATGTAAG CGTAGCTACG ACGAAACGTC AAAACCTTCT GGTAGACTCG TA - #CCTTAAAG        2640                                                                           - GTGTGCGCGA CGATGTTGCG TTTGTAGACC ACCATGATGC CCTCGTCCAG GT - #CTTCATTG        2700                                                                           - ATGGGCTTCA TCGAGGTGCA GACGATATTA CGTTCAAAGC GAATAAGATC CG - #TACCCTGA        2760                                                                           - GCCATAGAAC ACACGCGATA GGGGTACTTG GTGGTGTTGA CCCCCACCAC AT - #CTCCGTAC        2820                                                                           - TTGAGGGTAG TGTTGTAGAT GGTCTCGTTA ACACCATGGC TGACCGTTTG GG - #AAGAAGTT        2880                                                                           - ACGCGTTGAG AGACTGAACC GGATCGAGAA TGAGCAGCAG ACGTCGTATG AG - #AGGAATGG        2940                                                                           - TGACTGTGAG TAGCAGAAGT TCCACGAGTA GAAGATGAGG AAACCGCAGC AC - #CCAGACAG        3000                                                                           - ACGATACACA AGTTAACGCA GACTACCAGG CACCAGATCC TGGATTCCAT TA - #CGATACAA        3060                                                                           - ACTTAACGGA TATCGCGATA ATGAAATAAT TTATGATTAT TTCTCGCTTT CA - #ATTTAACA        3120                                                                           - CAACCCTCAA GAACCTTTGT ATTTATTTTC ACTTTTTAAG TATAGAATAA AG - #AAGCTGGG        3180                                                                           - AATCGATTCG CGATAGCTGA TTAGTTTTTG TTAACAAAAA TGTGGGAGAA TC - #TAATTAGT        3240                                                                           - TTTTCTTTAC ACAATTGACG TACATGAGTC TGAGTTCCTT GTTTTTGCTA AT - #TATTTCAT        3300                                                                           - CCAATTTATT ATTCTTGACG ATATCGAGAT CTTTTGTATA GGAGTCAGAC TT - #GTATTCAA        3360                                                                           - CATGCTTTTC TATAATCATC TTAGTTATTT CGGCATCATC CAATAGTACA TT - #TTCCAGAT        3420                                                                           - TAACAGAGTA GATATTAATG TCGTATTTGA ACAGAGCCTG TAACATCTCA AT - #GTCTTTAT        3480                                                                           - TATCTATAGC CAATTTAATG TCCGGAATGA AGAGAAGGGA ATTATTGGTG TT - #TGTCGACG        3540                                                                           - TCATATAGTC GAGCAAGAGA ATCATCATAT CCACGTGTCC ATTTTTTATA GT - #GGTGTGAA        3600                                                                           - TACAACTAAG GAGAATAGCC AGATCAAAAG TAGATGGTAT TTCTGAAAGA AA - #GTATGATA        3660                                                                           - CAATACTTAC ATCATTAAGC ATGACGGCAT GATAAAATGA AGTTTTCCAT CC - #AGTTTTCC        3720                                                                           - CATAGAACAT CAGTCTCCAA TTTTTCTTAA ACAGTTTCAC CGTTTGCATG TT - #ACCACTAT        3780                                                                           - CAACCGCATA ATACAATGCG GTGTTTCCTT TGTCATCAAA TTGTGAATCA TC - #CATTCCAC        3840                                                                           - TGAATAGCAA AATCTTTACT ATTTTGGTAT CTTCTAATGT GGCTGCCTGA TG - #TAATGGAA        3900                                                                           - ATTCATTCTC TAGAAGATTT TTCAATGCTC CAGCGTTCAA CAACGTACAT AC - #TAGACGCA        3960                                                                           - CGTTATTATC AGCTATTGCA TAATACAAGG CACTATGTCC ATGGACATCC GC - #CTTAAATG        4020                                                                           - TATCTTTACT AGAGAGAAAG CTTTTCAGCT GCTTAGACTT CCAAGTATTA AT - #TCGTGACA        4080                                                                           - GATCCATGTC TGAAACGAGA CGCTAATTAG TGTATATTTT TTCATTTTTT AT - #AATTTTGT        4140                                                                           - CATATTGCAC CAGAATTAAT AATATCTCTA ATAGATCTAA TTTAATTTAA TT - #TATATAAC        4200                                                                           - TTATTTTTTG AATATACTTT TAATTAACAA AAGAGTTAAG TTACTCATAT GG - #ACGCCGTC        4260                                                                           - CAGTCTGAAC ATCAATCTTT TTAGCCAGAG ATATCATAGC CGCTCTTAGA GT - #TTCAGCGT        4320                                                                           - GATTTTCCAA CCTAAATAGA ACTTCATCGT TGCGTTTACA ACACTTTTCT AT - #TTGTTCAA        4380                                                                           - ACTTTGTTGT TACATTAGTA ATCTTTTTTT CCAAATTAGT TAGCCGTTGT TT - #GAGAGTTT        4440                                                                           - CCTCATTGTC GTCTTCATCG GCTTTAACAA TTGCTTCGCG TTTAGCCTCC TG - #GCTGTTCT        4500                                                                           - TATCAGCCTT TGTAGAAAAA AATTCAGTTG CTGGAATTGC AAGATCGTCA TC - #TCCGGGGA        4560                                                                           #      4594        TTAA AGCCGCGGGA ATTC                                        - (2) INFORMATION FOR SEQ ID NO:44:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 2550 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:44:                                 - ATGGAATCCA GGATCTGGTG CCTGGTAGTC TGCGTTAACT TGTGTATCGT CT - #GTCTGGGT          60                                                                           - GCTGCGGTTT CCTCATCTTC TACTCGTGGA ACTTCTGCTA CTCACAGTCA CC - #ATTCCTCT         120                                                                           - CATACGACGT CTGCTGCTCA TTCTCGATCC GGTTCAGTCT CTCAACGCGT AA - #CTTCTTCC         180                                                                           - CAAACGGTCA GCCATGGTGT TAACGAGACC ATCTACAACA CTACCCTCAA GT - #ACGGAGAT         240                                                                           - GTGGTGGGGG TCAACACCAC CAAGTACCCC TATCGCGTGT GTTCTATGGC TC - #AGGGTACG         300                                                                           - GATCTTATTC GCTTTGAACG TAATATCGTC TGCACCTCGA TGAAGCCCAT CA - #ATGAAGAC         360                                                                           - CTGGACGAGG GCATCATGGT GGTCTACAAA CGCAACATCG TCGCGCACAC CT - #TTAAGGTA         420                                                                           - CGAGTCTACC AGAAGGTTTT GACGTTTCGT CGTAGCTACG CTTACATCCA CA - #CCACTTAT         480                                                                           - CTGCTGGGCA GCAACACGGA ATACGTGGCG CCTCCTATGT GGGAGATTCA TC - #ATATCAAC         540                                                                           - AGTCACAGTC AGTGCTACAG TTCCTACAGC CGCGTTATAG CAGGCACGGT TT - #TCGTGGCT         600                                                                           - TATCATAGGG ACAGCTATGA AAACAAAACC ATGCAATTAA TGCCCGACGA TT - #ATTCCAAC         660                                                                           - ACCCACAGTA CCCGTTACGT GACGGTCAAG GATCAATGGC ACAGCCGCGG CA - #GCACCTGG         720                                                                           - CTCTATCGTG AGACCTGTAA TCTGAATTGT ATGGTGACCA TCACTACTGC GC - #GCTCCAAG         780                                                                           - TATCCCTATC ATTTTTTCGC AACTTCCACG GGTGATGTGG TTGACATTTC TC - #CTTTCTAC         840                                                                           - AACGGAACTA ATCGCAATGC CAGCTATTTT GGAGAAAACG CCGACAAGTT TT - #TCATTTTT         900                                                                           - CCGAACTACA CTATCGTCTC CGACTTTGAA AGACCGAATT CTGCGTTAGA GA - #CCCACAGG         960                                                                           - TTGGTGGCTT TTCTTGAACG TGCGGACTCA GTGATCTCCT GGGATATACA GG - #ACGAGAAG        1020                                                                           - AATGTTACTT GTCAACTCAC TTTCTGGGAA GCCTCGGAAC GCACCATTCG TT - #CCGAAGCC        1080                                                                           - GAGGACTCGT ATCACTTTTC TTCTGCCAAA ATGACCGCCA CTTTCTTATC TA - #AGAAGCAA        1140                                                                           - GAGGTGAACA TGTCCGACTC TGCGCTGGAC TGTGTACGTG ATGAGGCCAT AA - #ATAAGTTA        1200                                                                           - CAGCAGATTT TCAATACTTC ATACAATCAA ACATATGAAA AATATGGAAA CG - #TGTCCGTC        1260                                                                           - TTTGAAACCA CTGGTGGTTT GGTGGTGTTC TGGCAAGGTA TCAAGCAAAA AT - #CTCTGGTG        1320                                                                           - GAACTCGAAC GTTTGGCCAA CCGCTCCAGT CTGAATCTTA CTCATAATAG AA - #CCATAAGA        1380                                                                           - TCTACAGATG GCAACAATGC AACTCATTTA TCCAACATGG AGTCGGTGCA CA - #ATCTGGTC        1440                                                                           - TACGCCCAGC TGCAGTTCAC CTATGACACG TTGCGCGGTT ACATCAACCG GG - #CGCTGGCC        1500                                                                           - GAAATCGCAG AAGCCTGGTG TGTGGATCAA CGGCGCACCC TAGAGGTCTT CA - #AGGAACTT        1560                                                                           - AGCAAGATCA ACCCGTCAGC TATTCTCTCG GCCATCTACA ACAAACCGAT TG - #CCGCGCGT        1620                                                                           - TTCATGGGTG ATGTCCTGGG TCTGGCCAGC TGCGTGACCA TTAACCAAAC CA - #GCGTCAAG        1680                                                                           - GTGCTGCGTG ATATGAATGT GAAGGAATCG CCAGGACGCT GCTACTCACG AC - #CAGTGGTC        1740                                                                           - ATCTTTAATT TCGCCAACAG CTCGTACGTG CAGTACGGTC AACTGGGCGA GG - #ATAACGAA        1800                                                                           - ATCCTGTTGG GCAACCACCG CACTGAGGAA TGTCAGCTTC CCAGCCTCAA GA - #TCTTCATC        1860                                                                           - GCCGGCAACT CGGCCTACGA GTACGTGGAC TACCTCTTCA AACGCATGAT TG - #ACCTCAGC        1920                                                                           - AGCATCTCCA CCGTCGACAG CATGATCGCC CTAGACATCG ACCCGCTGGA AA - #ACACCGAC        1980                                                                           - TTCAGGGTAC TGGAACTTTA CTCGCAGAAA GAATTGCGTT CCAGCAACGT TT - #TTGATCTC        2040                                                                           - GAGGAGATCA TGCGCGAGTT CAATTCGTAT AAGCAGCGGG TAAAGTACGT GG - #AGGACAAG        2100                                                                           - GTAGTCGACC CGCTGCCGCC CTACCTCAAG GGTCTGGACG ACACTCGACA GC - #GGCGTCTC        2160                                                                           - TGCATGCAGC CGCTGCAGAA CCTCTTTCCC TATCTGGTGT CCGCCGACGG GA - #CCACCGTG        2220                                                                           - ACGTCGGGCA ACACCAAAGA CACGTCGTTA CAGGCTCCGC CTTCCTACGA GG - #AAAGTGTT        2280                                                                           - TATAATTCTG GTCGCAAAGG ACCGGGACCA CCGTCGTCTG ATGCATCCAC GG - #CGGCTCCG        2340                                                                           - CCTTACACCA ACGAGCAGGC TTACCAGATG CTTCTGGCCC TGGTCCGTCT GG - #ACGCAGAG        2400                                                                           - CAGCGAGCGC ACGAGAACGG TACAGATTCT TTGGACGGAC AGACTGGCAC GC - #AGGACAAG        2460                                                                           - GGACAGAAGC CCAACCTGCT AGACCGACTG CGACACCGCA AAAACGGCTA CC - #GACACTTG        2520                                                                           #         2550     AAGA GAACGTCTGA                                             - (2) INFORMATION FOR SEQ ID NO:45:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 4594 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:45:                                 - AAGCTTGCGG CCGCTCATTA GACAAGCGAA TGAGGGACGA AAACGTGGAG GA - #GGTATTAA          60                                                                           - GTTTGGAGAA ATGGAGAGAG ACTGTTTAAT AGCGCATGGC GCAGCCAATA CT - #ATTACAGA         120                                                                           - AGTTTTGAAA GATTCGGAAG AAGATTATCA AGATGTGTAT GTTTGTGAAA AT - #TGTGGAGA         180                                                                           - CATAGCAGCA CAAATCAAGG GTATTAATAC ATGTCTTAGA TGTTCAAAAC TT - #AATCTCTC         240                                                                           - TCCTCTCTTA ACAAAAATTG ATACCACGCA CGTATCTAAA GTATTTCTTA CT - #CAAATGAA         300                                                                           - CGCCAGAGGC GTAAAAGTCA AATTAGATTT CGAACGAAGG CCTCCTTCGT TT - #TATAAACC         360                                                                           - ATTAGATAAA GTTGATCTCA AGCCGTCTTT TCTGGTGTAA TAAAAATTAA TT - #AATTACTC         420                                                                           - GAGGGTACCG GATCCCCCAG CTTATAAAAA TCACAAGTCT CTGACACTTT TT - #TTGTCTAG         480                                                                           - TTTTTTTTTC TCCTCTTGGT TCAGACGTTC TCTTCTTCGT CGGAGTCTTT CA - #AGTGTCGG         540                                                                           - TAGCCGTTTT TGCGGTGTCG CAGTCGGTCT AGCAGGTTGG GCTTCTGTCC CT - #TGTCCTGC         600                                                                           - GTGCCAGTCT GTCCGTCCAA AGAATCTGTA CCGTTCTCGT GCGCTCGCTG CT - #CTGCGTCC         660                                                                           - AGACGGACCA GGGCCAGAAG CATCTGGTAA GCCTGCTCGT TGGTGTAAGG CG - #GAGCCGCC         720                                                                           - GTGGATGCAT CAGACGACGG TGGTCCCGGT CCTTTGCGAC CAGAATTATA AA - #CACTTTCC         780                                                                           - TCGTAGGAAG GCGGAGCCTG TAACGACGTG TCTTTGGTGT TGCCCGACGT CA - #CGGTGGTC         840                                                                           - CCGTCGGCGG ACACCAGATA GGGAAAGAGG TTCTGCAGCG GCTGCATGCA GA - #GACGCCGC         900                                                                           - TGTCGAGTGT CGTCCAGACC CTTGAGGTAG GGCGGCAGCG GGTCGACTAC CT - #TGTCCTCC         960                                                                           - ACGTACTTTA CCCGCTGCTT ATACGAATTG AACTCGCGCA TGATCTCCTC GA - #GATCAAAA        1020                                                                           - ACGTTGCTGG AACGCAATTC TTTCTGCGAG TAAAGTTCCA GTACCCTGAA GT - #CGGTGTTT        1080                                                                           - TCCAGCGGGT CGATGTCTAG GGCGATCATG CTGTCGACGG TGGAGATGCT GC - #TGAGGTCA        1140                                                                           - ATCATGCGTT TGAAGAGGTA GTCCACGTAC TCGTAGGCCG AGTTGCCGGC GA - #TGAAGATC        1200                                                                           - TTGAGGCTGG GAAGCTGACA TTCCTCAGTG CGGTGGTTGC CCAACAGGAT TT - #CGTTATCC        1260                                                                           - TCGCCCAGTT GACCGTACTG CACGTACGAG CTGTTGGCGA AATTAAAGAT GA - #CCACTGGT        1320                                                                           - CGTGAGTAGC AGCGTCCTGG CGATTCCTTC ACATTCATAT CACGCAGCAC CT - #TGACGCTG        1380                                                                           - GTTTGGTTAA TGGTCACGCA GCTGGCCAGA CCCAGGACAT CACCCATGAA AC - #GCGCGGCA        1440                                                                           - ATCGGTTTGT TGTAGATGGC CGAGAGAATA GCTGACGGGT TGATCTTGCT AA - #GTTCCTTG        1500                                                                           - AAGACCTCTA GGGTGCGCCG TTGATCCACA CACCAGGCTT CTGCGATTTC GG - #CCAGCGCC        1560                                                                           - CGGTTGATGT AACCGCGCAA CGTGTCATAG GTGAACTGCA GCTGGGCGTA GA - #CCAGATTG        1620                                                                           - TGCACCGACT CCATGTTGGA TAAATGAGTT GCATTGTTGC CATCTGTAGA TC - #TTATGGTT        1680                                                                           - CTATTATGAG TAAGATTCAG ACTGGAGCGG TTGGCCAAAC GTTCGAGTTC CA - #CCAGAGAT        1740                                                                           - TTTTGCTTGA TACCTTGCCA GAACACCACC AAACCACCAG TGGTTTCAAA GA - #CGGACACG        1800                                                                           - TTTCCATATT TTTCATATGT TTGATTGTAT GAAGTATTGA AAATCTGCTG TA - #ACTTATTT        1860                                                                           - ATGGCCTCAT CACGTACACA GTCCAGCGCA GAGTCGGACA TGTTCACCTC TT - #GCTTCTTA        1920                                                                           - GATAAGAAAG TGGCGGTCAT TTTGGCAGAA GAAAAGTGAT ACGAGTCCTC GG - #CTTCGGAA        1980                                                                           - CGAATGGTGC GTTCCGAGGC TTCCCAGAAA GTGAGTTGAC AAGTAACATT CT - #TCTCGTCC        2040                                                                           - TGTATATCCC AGGAGATCAC TGAGTCCGCA CGTTCAAGAA AAGCCACCAA CC - #TGTGGGTC        2100                                                                           - TCTAACGCAG AATTCGGTCT TTCAAAGTCG GAGACGATAG TGTAGTTCGG AA - #AAATGAAA        2160                                                                           - AACTTGTCGG CGTTTTCTCC AAAATAGCTG GCATTGCGAT TAGTTCCGTT GT - #AGAAAGGA        2220                                                                           - GAAATGTCAA CCACATCACC CGTGGAAGTT GCGAAAAAAT GATAGGGATA CT - #TGGAGCGC        2280                                                                           - GCAGTAGTGA TGGTCACCAT ACAATTCAGA TTACAGGTCT CACGATAGAG CC - #AGGTGCTG        2340                                                                           - CCGCGGCTGT GCCATTGATC CTTGACCGTC ACGTAACGGG TACTGTGGGT GT - #TGGAATAA        2400                                                                           - TCGTCGGGCA TTAATTGCAT GGTTTTGTTT TCATAGCTGT CCCTATGATA AG - #CCACGAAA        2460                                                                           - ACCGTGCCTG CTATAACGCG GCTGTAGGAA CTGTAGCACT GACTGTGACT GT - #TGATATGA        2520                                                                           - TGAATCTCCC ACATAGGAGG CGCCACGTAT TCCGTGTTGC TGCCCAGCAG AT - #AAGTGGTG        2580                                                                           - TGGATGTAAG CGTAGCTACG ACGAAACGTC AAAACCTTCT GGTAGACTCG TA - #CCTTAAAG        2640                                                                           - GTGTGCGCGA CGATGTTGCG TTTGTAGACC ACCATGATGC CCTCGTCCAG GT - #CTTCATTG        2700                                                                           - ATGGGCTTCA TCGAGGTGCA GACGATATTA CGTTCAAAGC GAATAAGATC CG - #TACCCTGA        2760                                                                           - GCCATAGAAC ACACGCGATA GGGGTACTTG GTGGTGTTGA CCCCCACCAC AT - #CTCCGTAC        2820                                                                           - TTGAGGGTAG TGTTGTAGAT GGTCTCGTTA ACACCATGGC TGACCGTTTG GG - #AAGAAGTT        2880                                                                           - ACGCGTTGAG AGACTGAACC GGATCGAGAA TGAGCAGCAG ACGTCGTATG AG - #AGGAATGG        2940                                                                           - TGACTGTGAG TAGCAGAAGT TCCACGAGTA GAAGATGAGG AAACCGCAGC AC - #CCAGACAG        3000                                                                           - ACGATACACA AGTTAACGCA GACTACCAGG CACCAGATCC TGGATTCCAT TA - #CGATACAA        3060                                                                           - ACTTAACGGA TATCGCGATA ATGAAATAAT TTATGATTAT TTCTCGCTTT CA - #ATTTAACA        3120                                                                           - CAACCCTCAA GAACCTTTGT ATTTATTTTC ACTTTTTAAG TATAGAATAA AG - #AAGCTGGG        3180                                                                           - AATCGATTCG CGATAGCTGA TTAGTTTTTG TTAACAAAAA TGTGGGAGAA TC - #TAATTAGT        3240                                                                           - TTTTCTTTAC ACAATTGACG TACATGAGTC TGAGTTCCTT GTTTTTGCTA AT - #TATTTCAT        3300                                                                           - CCAATTTATT ATTCTTGACG ATATCGAGAT CTTTTGTATA GGAGTCAGAC TT - #GTATTCAA        3360                                                                           - CATGCTTTTC TATAATCATC TTAGTTATTT CGGCATCATC CAATAGTACA TT - #TTCCAGAT        3420                                                                           - TAACAGAGTA GATATTAATG TCGTATTTGA ACAGAGCCTG TAACATCTCA AT - #GTCTTTAT        3480                                                                           - TATCTATAGC CAATTTAATG TCCGGAATGA AGAGAAGGGA ATTATTGGTG TT - #TGTCGACG        3540                                                                           - TCATATAGTC GAGCAAGAGA ATCATCATAT CCACGTGTCC ATTTTTTATA GT - #GGTGTGAA        3600                                                                           - TACAACTAAG GAGAATAGCC AGATCAAAAG TAGATGGTAT TTCTGAAAGA AA - #GTATGATA        3660                                                                           - CAATACTTAC ATCATTAAGC ATGACGGCAT GATAAAATGA AGTTTTCCAT CC - #AGTTTTCC        3720                                                                           - CATAGAACAT CAGTCTCCAA TTTTTCTTAA ACAGTTTCAC CGTTTGCATG TT - #ACCACTAT        3780                                                                           - CAACCGCATA ATACAATGCG GTGTTTCCTT TGTCATCAAA TTGTGAATCA TC - #CATTCCAC        3840                                                                           - TGAATAGCAA AATCTTTACT ATTTTGGTAT CTTCTAATGT GGCTGCCTGA TG - #TAATGGAA        3900                                                                           - ATTCATTCTC TAGAAGATTT TTCAATGCTC CAGCGTTCAA CAACGTACAT AC - #TAGACGCA        3960                                                                           - CGTTATTATC AGCTATTGCA TAATACAAGG CACTATGTCC ATGGACATCC GC - #CTTAAATG        4020                                                                           - TATCTTTACT AGAGAGAAAG CTTTTCAGCT GCTTAGACTT CCAAGTATTA AT - #TCGTGACA        4080                                                                           - GATCCATGTC TGAAACGAGA CGCTAATTAG TGTATATTTT TTCATTTTTT AT - #AATTTTGT        4140                                                                           - CATATTGCAC CAGAATTAAT AATATCTCTA ATAGATCTAA TTTAATTTAA TT - #TATATAAC        4200                                                                           - TTATTTTTTG AATATACTTT TAATTAACAA AAGAGTTAAG TTACTCATAT GG - #ACGCCGTC        4260                                                                           - CAGTCTGAAC ATCAATCTTT TTAGCCAGAG ATATCATAGC CGCTCTTAGA GT - #TTCAGCGT        4320                                                                           - GATTTTCCAA CCTAAATAGA ACTTCATCGT TGCGTTTACA ACACTTTTCT AT - #TTGTTCAA        4380                                                                           - ACTTTGTTGT TACATTAGTA ATCTTTTTTT CCAAATTAGT TAGCCGTTGT TT - #GAGAGTTT        4440                                                                           - CCTCATTGTC GTCTTCATCG GCTTTAACAA TTGCTTCGCG TTTAGCCTCC TG - #GCTGTTCT        4500                                                                           - TATCAGCCTT TGTAGAAAAA AATTCAGTTG CTGGAATTGC AAGATCGTCA TC - #TCCGGGGA        4560                                                                           #      4594        TTAA AGCCGCGGGA ATTC                                        - (2) INFORMATION FOR SEQ ID NO:46:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 2229 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46:                                 - ATGCGGCCAG GCCTCCCCTC CTACCTCATC GTCCTCGCCG TCTGTCTCCT CA - #GCCACCTA          60                                                                           - CTTTCGTCAC GATATGGCGC AGAAGCCATA TCCGAACCGC TGGACAAAGC GT - #TTCACCTA         120                                                                           - CTGCTCAACA CCTACGGGAG ACCCATCCGC TTCCTGCGTG AAAACACCAC CC - #AGTGTACC         180                                                                           - TACAATAGCA GCCTCCGTAA CAGCACGGTC GTCAGGGAAA ACGCCATCAG TT - #TCAACTTT         240                                                                           - TTCCAAAGCT ATAATCAATA CTATGTATTC CATATGCCTC GATGTCTTTT TG - #CGGGTCCT         300                                                                           - CTGGCGGAGC AGTTTCTGAA CCAGGTAGAT CTGACCGAAA CCCTGGAAAG AT - #ACCAACAG         360                                                                           - AGACTTAACA CTTACGCGCT GGTATCCAAA GACCTGGCCA GCTACCGATC TT - #TTTCGCAG         420                                                                           - CAGCTAAAGG CACAGGACAG CCTAGGTGAA CAGCCCACCA CTGTGCCACC AC - #CCATTGAC         480                                                                           - CTGTCAATAC CTCACGTTTG GATGCCACCG CAAACCACTC CACACGGCTG GA - #CAGAATCA         540                                                                           - CATACCACCT CAGGACTACA CCGACCACAC TTTAACCAGA CCTGTATCCT CT - #TTGATGGA         600                                                                           - CACGATCTAC TATTCAGCAC CGTCACACCT TGTTTGCACC AAGGCTTTTA CC - #TCATCGAC         660                                                                           - GAACTACGTT ACGTTAAAAT AACACTGACC GAGGACTTCT TCGTAGTTAC GG - #TGTCCATA         720                                                                           - GACGACGACA CACCCATGCT GCTTATCTTC GGCCATCTTC CACGCGTACT CT - #TTAAAGCG         780                                                                           - CCCTATCAAC GCGACAACTT TATACTACGA CAAACTGAAA AACACGAGCT CC - #TGGTGCTA         840                                                                           - GTTAAGAAAG ATCAACTGAA CCGTCACTCT TATCTCAAAG ACCCGGACTT TC - #TTGACGCC         900                                                                           - GCACTTGACT TCAACTACCT GGACCTCAGC GCACTACTAC GTAACAGCTT TC - #ACCGTTAC         960                                                                           - GCCGTGGATG TACTCAAAAG CGGTCGATGT CAGATGCTGG ACCGCCGCAC GG - #TAGAAATG        1020                                                                           - GCCTTCGCCT ACGCATTAGC ACTGTTCGCA GCAGCCCGAC AAGAAGAGGC CG - #GCGCCCAA        1080                                                                           - GTCTCCGTCC CACGGGCCCT AGACCGCCAG GCCGCACTCT TACAAATACA AG - #AATTTATG        1140                                                                           - ATCACCTGCC TCTCACAAAC ACCACCACGC ACCACGTTGC TGCTGTATCC CA - #CGGCCGTG        1200                                                                           - GACCTGGCCA AACGAGCCCT TTGGACACCG AATCAGATCA CCGACATCAC CA - #GCCTCGTA        1260                                                                           - CGCCTGGTCT ACATACTCTC TAAACAGAAT CAGCAACATC TCATCCCCCA GT - #GGGCACTA        1320                                                                           - CGACAGATCG CCGACTTTGC CCTAAAACTA CACAAAACGC ACCTGGCCTC TT - #TTCTTTCA        1380                                                                           - GCCTTCGCGC GTCAAGAACT CTACCTCATG GGCAGCCTCG TCCACTCCAT GC - #TAGTACAT        1440                                                                           - ACGACGGAGA GACGCGAAAT CTTCATCGTA GAAACGGGCC TCTGTTCATT AG - #CCGAGCTA        1500                                                                           - TCACACTTTA CGCAGTTGCT AGCTCATCCG CACCACGAAT ACCTCAGCGA CC - #TGTACACA        1560                                                                           - CCCTGTTCCA GTAGCGGGCG ACGCGATCAC TCGCTCGAAC GCCTCACACG TC - #TCTTCCCC        1620                                                                           - GATGCCACCG TCCCCACTAC CGTTCCCGCC GCCCTCTCCA TCCTATCTAC CA - #TGCAACCA        1680                                                                           - AGCACGCTAG AAACCTTCCC CGACCTGTTT TGTCTGCCGC TCGGCGAATC CT - #TCTCCGCG        1740                                                                           - CTGACCGTCT CCGAACACGT CAGTTATGTC GTAACAAACC AGTACCTGAT CA - #AAGGTATC        1800                                                                           - TCCTACCCTG TCTCCACCAC CGTCGTAGGC CAGAGCCTCA TCATCACCCA GA - #CGGACAGT        1860                                                                           - CAAACTAAAT GCGAACTGAC GCGCAACATG CATACCACAC ACAGCATCAC AG - #CGGCGCTC        1920                                                                           - AACATTTCCC TAGAAAACTG CGCCTTTTGC CAAAGCGCCC TACTAGAATA CG - #ACGACACG        1980                                                                           - CAAGGCGTCA TCAACATCAT GTACATGCAC GACTCGGACG ACGTCCTTTT CG - #CCCTGGAT        2040                                                                           - CCCTACAACG AAGTGGTGGT CTCATCTCCG CGAACTCACT ACCTCATGCT TT - #TGAAAAAC        2100                                                                           - GGTACGGTCC TAGAAGTAAC TGACGTCGTC GTGGACGCTA CCGACAGTCG TC - #TCCTCATG        2160                                                                           - ATGTCCGTCT ACGCGCTATC GGCCATCATC GGCATCTATC TGCTCTACCG CA - #TGCTCAAG        2220                                                                           #       2229                                                                   - (2) INFORMATION FOR SEQ ID NO:47:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 3539 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:47:                                 - CTGCAGGTCG ACGGATCTGA GAATGGATGA TTCTCCAGCC GAAACATATT CT - #ACCATGGC          60                                                                           - TCCGTTTAAT TTGTTGATGA AGATGGATTC ATCCTTAAAT GTTTTCTCTG TA - #ATAGTTTC         120                                                                           - CACCGAAAGA CTATGCAAAG AATTTGGAAT GCGTTCCTTG TGCTTAATGT TT - #CCATAGAC         180                                                                           - GGCTTCTAGA AGTTGATACA ACATAGGACT AGCCGCGGTA ACTTTTATTT TT - #AGAAAGTA         240                                                                           - TCCATCGCTT CTATCTTGTT TAGATTTATT TTTATAAAGT TTAGTCTCTC CT - #TCCAACAT         300                                                                           - AATAAAAGTG GAAGTCATTT GACTAGATAA ACTATCAGTA AGTTTTATAG AG - #ATAGACGA         360                                                                           - ACAATTAGCG TATTGAGAAG CATTTAGTGT AACGTATTCG ATACATTTTG CA - #TTAGATTT         420                                                                           - ACTAATCGAT TTTGCATACT CTATAACACC CGCACAAGTC TGTAGAGAAT CG - #CTAGATGC         480                                                                           - AGTAGGTCTT GGTGAAGTTT CAACTCTCTT CTTGATTACC TTACTCATGA TT - #AAACCTAA         540                                                                           - ATAATTGTAC TTTGTAATAT AATGATATAT ATTTTCACTT TATCTCATTT GA - #GAATAAAA         600                                                                           - AGATCACAAA AATTAACTAA TCAGGATCCG GTACCCTCGA GTTTATTGGG AA - #GAATATGA         660                                                                           - TAATATTTTG GGATTTCAAA ATTGAAAATA TATAATTACA ATATAAAATG CG - #GCCCGGGC         720                                                                           - TCCCCTCCTA CCTCATCGTC CTCGCCGTCT GTCTCCTCAG CCACCTACTT TC - #GTCACGAT         780                                                                           - ATGGCGCAGA AGCCATATCC GAACCGCTGG ACAAAGCGTT TCACCTACTG CT - #CAACACCT         840                                                                           - ACGGGAGACC CATCCGCTTC CTGCGTGAAA ACACCACCCA GTGTACCTAC AA - #TAGCAGCC         900                                                                           - TCCGTAACAG CACGGTCGTC AGGGAAAACG CCATCAGTTT CAACTTTTTC CA - #AAGCTATA         960                                                                           - ATCAATACTA TGTATTCCAT ATGCCTCGAT GTCTTTTTGC GGGTCCTCTG GC - #GGAGCAGT        1020                                                                           - TTCTGAACCA GGTAGATCTG ACCGAAACCC TGGAAAGATA CCAACAGAGA CT - #TAACACTT        1080                                                                           - ACGCGCTGGT ATCCAAAGAC CTGGCCAGCT ACCGATCTTT TTCGCAGCAG CT - #AAAGGCAC        1140                                                                           - AGGACAGCCT AGGTGAACAG CCCACCACTG TGCCACCACC CATTGACCTG TC - #AATACCTC        1200                                                                           - ACGTTTGGAT GCCACCGCAA ACCACTCCAC ACGGCTGGAC AGAATCACAT AC - #CACCTCAG        1260                                                                           - GACTACACCG ACCACACTTT AACCAGACCT GTATCCTCTT TGATGGACAC GA - #TCTACTAT        1320                                                                           - TCAGCACCGT CACACCTTGT TTGCACCAAG GCTTTTACCT CATCGACGAA CT - #ACGTTACG        1380                                                                           - TTAAAATAAC ACTGACCGAG GACTTCTTCG TAGTTACGGT GTCCATAGAC GA - #CGACACAC        1440                                                                           - CCATGCTGCT TATCTTCGGC CATCTTCCAC GCGTACTCTT TAAAGCGCCC TA - #TCAACGCG        1500                                                                           - ACAACTTTAT ACTACGACAA ACTGAAAAAC ACGAGCTCCT GGTGCTAGTT AA - #GAAAGATC        1560                                                                           - AACTGAACCG TCACTCTTAT CTCAAAGACC CGGACTTTCT TGACGCCGCA CT - #TGACTTCA        1620                                                                           - ACTACCTGGA CCTCAGCGCA CTACTACGTA ACAGCTTTCA CCGTTACGCC GT - #GGATGTAC        1680                                                                           - TCAAAAGCGG TCGATGTCAG ATGCTGGACC GCCGCACGGT AGAAATGGCC TT - #CGCCTACG        1740                                                                           - CATTAGCACT GTTCGCAGCA GCCCGACAAG AAGAGGCCGG CGCCCAAGTC TC - #CGTCCCAC        1800                                                                           - GGGCCCTAGA CCGCCAGGCC GCACTCTTAC AAATACAAGA ATTTATGATC AC - #CTGCCTCT        1860                                                                           - CACAAACACC ACCACGCACC ACGTTGCTGC TGTATCCCAC GGCCGTGGAC CT - #GGCCAAAC        1920                                                                           - GAGCCCTTTG GACACCGAAT CAGATCACCG ACATCACCAG CCTCGTACGC CT - #GGTCTACA        1980                                                                           - TACTCTCTAA ACAGAATCAG CAACATCTCA TCCCCCAGTG GGCACTACGA CA - #GATCGCCG        2040                                                                           - ACTTTGCCCT AAAACTACAC AAAACGCACC TGGCCTCTTT TCTTTCAGCC TT - #CGCGCGTC        2100                                                                           - AAGAACTCTA CCTCATGGGC AGCCTCGTCC ACTCCATGCT AGTACATACG AC - #GGAGAGAC        2160                                                                           - GCGAAATCTT CATCGTAGAA ACGGGCCTCT GTTCATTAGC CGAGCTATCA CA - #CTTTACGC        2220                                                                           - AGTTGCTAGC TCATCCGCAC CACGAATACC TCAGCGACCT GTACACACCC TG - #TTCCAGTA        2280                                                                           - GCGGGCGACG CGATCACTCG CTCGAACGCC TCACACGTCT CTTCCCCGAT GC - #CACCGTCC        2340                                                                           - CCACTACCGT TCCCGCCGCC CTCTCCATCC TATCTACCAT GCAACCAAGC AC - #GCTAGAAA        2400                                                                           - CCTTCCCCGA CCTGTTTTGT CTGCCGCTCG GCGAATCCTT CTCCGCGCTG AC - #CGTCTCCG        2460                                                                           - AACACGTCAG TTATGTCGTA ACAAACCAGT ACCTGATCAA AGGTATCTCC TA - #CCCTGTCT        2520                                                                           - CCACCACCGT CGTAGGCCAG AGCCTCATCA TCACCCAGAC GGACAGTCAA AC - #TAAATGCG        2580                                                                           - AACTGACGCG CAACATGCAT ACCACACACA GCATCACAGC GGCGCTCAAC AT - #TTCCCTAG        2640                                                                           - AAAACTGCGC CTTTTGCCAA AGCGCCCTAC TAGAATACGA CGACACGCAA GG - #CGTCATCA        2700                                                                           - ACATCATGTA CATGCACGAC TCGGACGACG TCCTTTTCGC CCTGGATCCC TA - #CAACGAAG        2760                                                                           - TGGTGGTCTC ATCTCCGCGA ACTCACTACC TCATGCTTTT GAAAAACGGT AC - #GGTCCTAG        2820                                                                           - AAGTAACTGA CGTCGTCGTG GACGCTACCG ACAGTCGTCT CCTCATGATG TC - #CGTCTACG        2880                                                                           - CGCTATCGGC CATCATCGGC ATCTATCTGC TCTACCGCAT GCTCAAGACA TG - #CTGATTTT        2940                                                                           - TATCTCGAGC CCGGGAGATC TTAGCTAACT GATTTTTCTG GGAAAAAAAT TA - #TTTAACTT        3000                                                                           - TTCATTAATA GGGATTTGAC GTATGTAGCG TACAAAATTA TCGTTCCTGG TA - #TATAGATA        3060                                                                           - AAGAGTCCTA TATATTTGAA AATCGTTACG GCTCGATTAA ACTTTAATGA TT - #GCATAGTG        3120                                                                           - AATATATCAT TAGGATTTAA CTCCTTGACT ATCATGGCGG CGCCAGAAAT TA - #CCATCAAA        3180                                                                           - AGCATTAATA CAGTTATGCC GATCGCAGTT AGAACGGTTA TAGCATCCAC CA - #TTTATATC        3240                                                                           - TAAAAATTAG ATCAAAGAAT ATGTGACAAA GTCCTAGTTG TATACTGAGA AT - #TGACGAAA        3300                                                                           - CAATGTTTCT TACATATTTT TTTCTTATTA GTAACTGACT TAATAGTAGG AA - #CTGGAAAG        3360                                                                           - CTAGACTTGA TTATTCTATA AGTATAGATA CCCTTCCAGA TAATGTTCTC TT - #TGATAAAA        3420                                                                           - GTTCCAGAAA ATGTAGAATT TTTTAAAAAG TTATCTTTTG CTATTACCAA GA - #TTGTGTTT        3480                                                                           - AGACGCTTAT TATTAATATG AGTAATGAAA TCCACACCGC CTCTAGATAT GG - #GGAATTC         3539                                                                           - (2) INFORMATION FOR SEQ ID NO:48:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 4427 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:48:                                 - GAATTGCGGC CGCTGAATGT TAAATGTTAT ACTTTGGATG AAGCTATAAA TA - #TGCATTGG          60                                                                           - AAAAATAATC CATTTAAAGA AAGGATTCAA ATACTACAAA ACCTAAGCGA TA - #ATATGTTA         120                                                                           - ACTAAGCTTA TTCTTAACGA CGCTTTAAAT ATACACAAAT AAACATAATT TT - #TGTATAAC         180                                                                           - CTAACAAATA ACTAAAACAT AAAAATAATA AAAGGAAATG TAATATCGTA AT - #TATTTTAC         240                                                                           - TCAGGAATGG GGTTAAATAT TTATATCACG TGTATATCTA TACTGTTATC GT - #ATACTCTT         300                                                                           - TACAATTACT ATTACGAATA TGCAAGAGAT AATAAGATTA CGTATTTAAG AG - #AATCTTGT         360                                                                           - CATGATAATT GGGTACGACA TAGTGATAAA TGCTATTTCG CATCGTTACA TA - #AAGTCAGT         420                                                                           - TGGAAAGATG GATTTGACAG ATGTAACTTA ATAGGTGCAA AAATGTTAAA TA - #ACAGCATT         480                                                                           - CTATCGGAAG ATAGGATACC AGTTATATTA TACAAAAATC ACTGGTTGGA TA - #AAACAGAT         540                                                                           - TCTGCAATAT TCGTAAAAGA TGAAGATTAC TGCGAATTTG TAAACTATGA CA - #ATAAAAAG         600                                                                           - CCATTTATCT CAACGACATC GTGTAATTCT TCCATGTTTT ATGTATGTGT TT - #CAGATATT         660                                                                           - ATGAGATTAC TATAAACTTT TTGTATACTT ATATTCCGTA AACTATATTA AT - #CATGAAGA         720                                                                           - AAATGAAAAA GTATAGAAGC TGTTCACGAG CGGTTGTTGA AAACAACAAA AT - #TATACATT         780                                                                           - CAAGATGGCT TACATATACG TCTGTGAGGC TATCATGGAT AATGACAATG CA - #TCTCTAAA         840                                                                           - TAGGTTTTTG GACAATGGAT TCGACCCTAA CACGGAATAT GGTACTCTAC AA - #TCTCCTCT         900                                                                           - TGAAATGGCT GTAATGTTCA AGAATACCGA GGCTATAAAA ATCTTGATGA GG - #TATGGAGC         960                                                                           - TAAACCTGTA GTTACTGAAT GCACAACTTC TTGTCTGCAT GATGCGGTGT TG - #AGAGACGA        1020                                                                           - CTACAAAATA GTGAAAGATC TGTTGAAGAA TAACTATGTA AACAATGTTC TT - #TACAGCGG        1080                                                                           - AGGCTTTACT CCTTTGTGTT TGGCAGCTTA CCTTAACAAA GTTAATTTGG TT - #AAACTTCT        1140                                                                           - ATTGGCTCAT TCGGCGGATG TAGATATTTC AAACACGGAT CGGTTAACTC CT - #CTACATAT        1200                                                                           - AGCCGTATCA AATAAAAATT TAACAATGGT TAAACTTCTA TTGAACAAAG GT - #GCTGATAC        1260                                                                           - TGACTTGCTG GATAACATGG GACGTACTCC TTTAATGATC GCTGTACAAT CT - #GGAAATAT        1320                                                                           - TGAAATATGT AGCACACTAC TTAAAAAAAA TAAAATGTCC AGAACTGGGA AA - #AATTGATC        1380                                                                           - TTGCCAGCTG TAATTCATGG TAGAAAAGAA GTGCTCAGGC TACTTTTCAA CA - #AAGGAGCA        1440                                                                           - GATGTAAACT ACATCTTTGA AAGAAATGGA AAATCATATA CTGTTTTGGA AT - #TGATTAAA        1500                                                                           - GAAAGTTACT CTGAGACACA AAAGAGGTAG CTGAAGTGGT ACTCTCAAAG GT - #ACGTGACT        1560                                                                           - AATTAGCTAT AAAAAGGATC TTAATTAATT AGTCATCAGG CAGGGCGAGA AC - #GAGACTAT        1620                                                                           - CTGCTCGTTA ATTAATTAGG TCGACGGATC CGGTACCCTC GAGTTTATTG GG - #AAGAATAT        1680                                                                           - GATAATATTT TGGGATTTCA AAATTGAAAA TATATAATTA CAATATAAAA TG - #CGGCCCGG        1740                                                                           - GCTCCCCTCC TACCTCATCG TCCTCGCCGT CTGTCTCCTC AGCCACCTAC TT - #TCGTCACG        1800                                                                           - ATATGGCGCA GAAGCCATAT CCGAACCGCT GGACAAAGCG TTTCACCTAC TG - #CTCAACAC        1860                                                                           - CTACGGGAGA CCCATCCGCT TCCTGCGTGA AAACACCACC CAGTGTACCT AC - #AATAGCAG        1920                                                                           - CCTCCGTAAC AGCACGGTCG TCAGGGAAAA CGCCATCAGT TTCAACTTTT TC - #CAAAGCTA        1980                                                                           - TAATCAATAC TATGTATTCC ATATGCCTCG ATGTCTTTTT GCGGGTCCTC TG - #GCGGAGCA        2040                                                                           - GTTTCTGAAC CAGGTAGATC TGACCGAAAC CCTGGAAAGA TACCAACAGA GA - #CTTAACAC        2100                                                                           - TTACGCGCTG GTATCCAAAG ACCTGGCCAG CTACCGATCT TTTTCGCAGC AG - #CTAAAGGC        2160                                                                           - ACAGGACAGC CTAGGTGAAC AGCCCACCAC TGTGCCACCA CCCATTGACC TG - #TCAATACC        2220                                                                           - TCACGTTTGG ATGCCACCGC AAACCACTCC ACACGGCTGG ACAGAATCAC AT - #ACCACCTC        2280                                                                           - AGGACTACAC CGACCACACT TTAACCAGAC CTGTATCCTC TTTGATGGAC AC - #GATCTACT        2340                                                                           - ATTCAGCACC GTCACACCTT GTTTGCACCA AGGCTTTTAC CTCATCGACG AA - #CTACGTTA        2400                                                                           - CGTTAAAATA ACACTGACCG AGGACTTCTT CGTAGTTACG GTGTCCATAG AC - #GACGACAC        2460                                                                           - ACCCATGCTG CTTATCTTCG GCCATCTTCC ACGCGTACTC TTTAAAGCGC CC - #TATCAACG        2520                                                                           - CGACAACTTT ATACTACGAC AAACTGAAAA ACACGAGCTC CTGGTGCTAG TT - #AAGAAAGA        2580                                                                           - TCAACTGAAC CGTCACTCTT ATCTCAAAGA CCCGGACTTT CTTGACGCCG CA - #CTTGACTT        2640                                                                           - CAACTACCTG GACCTCAGCG CACTACTACG TAACAGCTTT CACCGTTACG CC - #GTGGATGT        2700                                                                           - ACTCAAAAGC GGTCGATGTC AGATGCTGGA CCGCCGCACG GTAGAAATGG CC - #TTCGCCTA        2760                                                                           - CGCATTAGCA CTGTTCGCAG CAGCCCGACA AGAAGAGGCC GGCGCCCAAG TC - #TCCGTCCC        2820                                                                           - ACGGGCCCTA GACCGCCAGG CCGCACTCTT ACAAATACAA GAATTTATGA TC - #ACCTGCCT        2880                                                                           - CTCACAAACA CCACCACGCA CCACGTTGCT GCTGTATCCC ACGGCCGTGG AC - #CTGGCCAA        2940                                                                           - ACGAGCCCTT TGGACACCGA ATCAGATCAC CGACATCACC AGCCTCGTAC GC - #CTGGTCTA        3000                                                                           - CATACTCTCT AAACAGAATC AGCAACATCT CATCCCCCAG TGGGCACTAC GA - #CAGATCGC        3060                                                                           - CGACTTTGCC CTAAAACTAC ACAAAACGCA CCTGGCCTCT TTTCTTTCAG CC - #TTCGCGCG        3120                                                                           - TCAAGAACTC TACCTCATGG GCAGCCTCGT CCACTCCATG CTAGTACATA CG - #ACGGAGAG        3180                                                                           - ACGCGAAATC TTCATCGTAG AAACGGGCCT CTGTTCATTA GCCGAGCTAT CA - #CACTTTAC        3240                                                                           - GCAGTTGCTA GCTCATCCGC ACCACGAATA CCTCAGCGAC CTGTACACAC CC - #TGTTCCAG        3300                                                                           - TAGCGGGCGA CGCGATCACT CGCTCGAACG CCTCACACGT CTCTTCCCCG AT - #GCCACCGT        3360                                                                           - CCCCACTACC GTTCCCGCCG CCCTCTCCAT CCTATCTACC ATGCAACCAA GC - #ACGCTAGA        3420                                                                           - AACCTTCCCC GACCTGTTTT GTCTGCCGCT CGGCGAATCC TTCTCCGCGC TG - #ACCGTCTC        3480                                                                           - CGAACACGTC AGTTATGTCG TAACAAACCA GTACCTGATC AAAGGTATCT CC - #TACCCTGT        3540                                                                           - CTCCACCACC GTCGTAGGCC AGAGCCTCAT CATCACCCAG ACGGACAGTC AA - #ACTAAATG        3600                                                                           - CGAACTGACG CGCAACATGC ATACCACACA CAGCATCACA GCGGCGCTCA AC - #ATTTCCCT        3660                                                                           - AGAAAACTGC GCCTTTTGCC AAAGCGCCCT ACTAGAATAC GACGACACGC AA - #GGCGTCAT        3720                                                                           - CAACATCATG TACATGCACG ACTCGGACGA CGTCCTTTTC GCCCTGGATC CC - #TACAACGA        3780                                                                           - AGTGGTGGTC TCATCTCCGC GAACTCACTA CCTCATGCTT TTGAAAAACG GT - #ACGGTCCT        3840                                                                           - AGAAGTAACT GACGTCGTCG TGGACGCTAC CGACAGTCGT CTCCTCATGA TG - #TCCGTCTA        3900                                                                           - CGCGCTATCG GCCATCATCG GCATCTATCT GCTCTACCGC ATGCTCAAGA CA - #TGCTGATT        3960                                                                           - TTTATCTCGA GTCTAGAATC GATCCCGGGT TTTTATGACT AGTTAATCAC GG - #CCGCTTAT        4020                                                                           - AAAGATCTAA AATGCATAAT TTCTAAATAA TGAAAAAAAA GTACATCATG AG - #CAACGCGT        4080                                                                           - TAGTATATTT TACAATGGAG ATTAACGCTC TATACCGTTC TATGTTTATT GA - #TTCAGATG        4140                                                                           - ATGTTTTAGA AAAGAAAGTT ATTGAATATG AAAACTTTAA TGAAGATGAA GA - #TGACGACG        4200                                                                           - ATGATTATTG TTGTAAATCT GTTTTAGATG AAGAAGATGA CGCGCTAAAG TA - #TACTATGG        4260                                                                           - TTACAAAGTA TAAGTCTATA CTACTAATGG CGACTTGTGC AAGAAGGTAT AG - #TATAGTGA        4320                                                                           - AAATGTTGTT AGATTATGAT TATGAAAAAC CAAATAAATC AGATCCATAT CT - #AAAGGTAT        4380                                                                           #              4427TTCA TCTATTCCTA GTTTAGAATA CCTGCAG                          - (2) INFORMATION FOR SEQ ID NO:49:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 2651 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49:                                 - AAGACTAATT TGTAAACCAT CTTACTCAAA ATATGTAACA ATAGTACGAT GC - #AATGAGTA          60                                                                           - AGACAATAGG AAATCTATCT TATATACACA TAATTATTCT ATCAATTTTA CC - #AATTAGTT         120                                                                           - AGTGTAATGT TATAAAAACT AATTAATCAC TCGAGATAAA AATCAGCATG TC - #TTGAGCAT         180                                                                           - GCGGTAGAGC AGATAGATGC CGATGATGGC CGATAGCGCG TAGACGGACA TC - #ATGAGGAG         240                                                                           - ACGACTGTCG GTAGCGTCCA CGACGACGTC AGTTACTTCT AGGACCGTAC CG - #TTTTTCAA         300                                                                           - AAGCATGAGG TAGTGAGTTC GCGGAGATGA GACCACCACT TCGTTGTAGG GA - #TCCAGGGC         360                                                                           - GAAAAGGACG TCGTCCGAGT CGTGCATGTA CATGATGTTG ATGACGCCTT GC - #GTGTCGTC         420                                                                           - GTATTCTAGT AGGGCGCTTT GGCAAAAGGC GCAGTTTTCT AGGGAAATGT TG - #AGCGCCGC         480                                                                           - TGTGATGCTG TGTGTGGTAT GCATGTTGCG CGTCAGTTCG CATTTAGTTT GA - #CTGTCCGT         540                                                                           - CTGGGTGATG ATGAGGCTCT GGCCTACGAC GGTGGTGGAG ACAGGGTAGG AG - #ATACCTTT         600                                                                           - GATCAGGTAC TGGTTTGTTA CGACATAACT GACGTGTTCG GAGACGGTCA GC - #GCGGAGAA         660                                                                           - GGATTCGCCG AGCGGCAGAC AAAACAGGTC GGGGAAGGTT TCTAGCGTGC TT - #GGTTGCAT         720                                                                           - GGTAGATAGG ATGGAGAGGG CGGCGGGAAC GGTAGTGGGG ACGGTGGCAT CG - #GGGAAGAG         780                                                                           - ACGTGTGAGG CGTTCGAGCG AGTGATCGCG TCGCCCGCTA CTGGAACAGG GT - #GTGTACAG         840                                                                           - GTCGCTGAGG TATTCGTGGT GCGGATGAGC TAGCAACTGC GTAAAGTGTG AT - #AGCTCGGC         900                                                                           - TAATGAACAG AGGCCCGTTT CTACGATGAA GATTTCGCGT CTCTCCGTCG TA - #TGTACTAG         960                                                                           - CATGGAGTGG ACGAGGCTGC CCATGAGGTA GAGTTCTTGA CGCGCGAAGG CT - #GAAAGAAA        1020                                                                           - AGAGGCCAGG TGCGTTTTGT GTAGTTTTAG GGCAAAGTCG GCGATCTGTC GT - #AGTGCCCA        1080                                                                           - CTGGGGGATG AGATGTTGCT GATTCTGTTT AGAGAGTATG TAGACCAGGC GT - #ACGAGGCT        1140                                                                           - GGTGATGTCG GTGATCTGAT TCGGTGTCCA AAGGGCTCGT TTGGCCAGGT CC - #ACGGCCGT        1200                                                                           - GGGATACAGC AGCAACGTGG TGCGTGGTGG TGTTTGTGAG AGGCAGGTGA TC - #ATAAATTC        1260                                                                           - TTGTATTTGT AAGAGTGCGG CCTGGCGGTC TAGGGCCCGT GGGACGGAGA CT - #TGGGCGCC        1320                                                                           - GGCCTCTTCT TGTCGGGCTG CTGCGAACAG TGCTAATGCG TAGGCGAAGG CC - #ATTTCTAC        1380                                                                           - CGTGCGGCGG TCCAGCATCT GACATCGACC GCTTTTGAGT ACATCCACGG CG - #TAACGGTG        1440                                                                           - AAAGCTGTTA CGTAGTAGTG CGCTGAGGTC CAGGTAGTTG AAGTCAAGTG CG - #GCGTCAAG        1500                                                                           - AAAGTCCGGG TCTTTGAGAT AAGAGTGACG GTTCAGTTGA TCTTTCTTAA CT - #AGCACCAG        1560                                                                           - GAGCTCGTGT TTTTCAGTTT GTCGTAGTAT AAAGTTGTCG CGTTGATAGG GC - #GCTTTAAA        1620                                                                           - GAGTACGCGT GGAAGATGGC CGAAGATAAG CAGCATGGGT GTGTCGTCGT CT - #ATGGACAC        1680                                                                           - CGTAACTACG AAGAAGTCCT CGGTCAGTGT TATTTTAACG TAACGTAGTT CG - #TCGATGAG        1740                                                                           - GTAAAAGCCT TGGTGCAAAC AAGGTGTGAC GGTGCTGAAT AGTAGATCGT GT - #CCATCAAA        1800                                                                           - GAGGATACAG GTCTGGTTAA AGTGTGGTCG GTGTAGTCCT GAGGTGGTAT GT - #GATTCTGT        1860                                                                           - CCAGCCGTGT GGAGTGGTTT GCGGTGGCAT CCAAACGTGA GGTATTGACA GG - #TCAATGGG        1920                                                                           - TGGTGGCACA GTGGTGGGCT GTTCACCTAG GCTGTCCTGT GCCTTTAGCT GC - #TGCGAAAA        1980                                                                           - AGATCGGTAG CTGGCCAGGT CTTTGGATAC CAGCGCGTAA GTGTTAAGTC TC - #TGTTGGTA        2040                                                                           - TCTTTCCAGG GTTTCGGTCA GATCTACCTG GTTCAGAAAC TGCTCCGCCA GA - #GGACCCGC        2100                                                                           - AAAAAGACAT CGAGGCATAT GGAATACATA GTATTGATTA TAGCTTTGGA AA - #AAGTTGAA        2160                                                                           - ACTGATGGCG TTTTCCCTGA CGACCGTGCT GTTACGGAGG CTGCTATTGT AG - #GTACACTG        2220                                                                           - GGTGGTGTTT TCACGCAGGA AGCGGATGGG TCTCCCGTAG GTGTTGAGCA GT - #AGGTGAAA        2280                                                                           - CGCTTTGTCC AGCGGTTCGG ATATGGCTTC TGCGCCATAT CGTGACGAAA GT - #AGGTGGCT        2340                                                                           - GAGGAGACAG ACGGCGAGGA CGATGAGGTA GGAGGGGAGC CCGGGCCGCA TT - #TTATATTG        2400                                                                           - TAATTATATA TTTTCAATTT TGAAATCCCA AAATATTATC ATATTCTTCC CA - #ATAAACTC        2460                                                                           - GAGCCCGGGG AATTCGGATC CTCGCGACTG CAGGGTACCT GAGTAGCTAA TT - #TTTAAACA        2520                                                                           - AAAATGTGGG AGAATCTAAT TAGTTTTTCT TTACACAATT GACGTACATG AG - #TCTGAGTT        2580                                                                           - CCTTGTTTTT GCTAATTATT TCATCCAATT TATTATTCTT GACGATATCG AG - #ATCTTTTG        2640                                                                           #     2651                                                                     - (2) INFORMATION FOR SEQ ID NO:50:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 1476 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50:                                 - ATGGAGTCCT CTGCCAAGAG AAAGATGGAC CCTGATAATC CTGACGAGGG CC - #CTTCCTCC          60                                                                           - AAGGTGCCAC GGCCCGAGAC ACCCGTGACC AAGGCCACGA CGTTCCTGCA GA - #CTATGTTG         120                                                                           - AGGAAGGAGG TTAACAGTCA GCTGAGTCTG GGAGACCCGC TGTTTCCAGA GT - #TGGCCGAA         180                                                                           - GAATCCCTCA AAACTTTTGA ACAAGTGACC GAGGATTGCA ACGAGAACCC CG - #AGAAAGAT         240                                                                           - GTCCTGGCAG AACTCGTCAA ACAGATTAAG GTTCGAGTGG ACATGGTGCG GC - #ATAGAATC         300                                                                           - AAGGAGCACA TGCTGAAAAA ATATACCCAG ACGGAAGAGA AATTCACTGG CG - #CCTTTAAT         360                                                                           - ATGATGGGAG GATGTTTGCA GAATGCCTTA GATATCTTAG ATAAGGTTCA TG - #AGCCTTTC         420                                                                           - GAGGAGATGA AGTGTATTGG GCTAACTATG CAGAGCATGT ATGAGAACTA CA - #TTGTACCT         480                                                                           - GAGGATAAGC GGGAGATGTG GATGGCTTGT ATTAAGGAGC TGCATGATGT GA - #GCAAGGGC         540                                                                           - GCCGCTAACA AGTTGGGGGG TGCACTGCAG GCTAAGGCCC GTGCTAAAAA GG - #ATGAACTT         600                                                                           - AGGAGAAAGA TGATGTATAT GTGCTACAGG AATATAGAGT TCTTTACCAA GA - #ACTCAGCC         660                                                                           - TTCCCTAAGA CCACCAATGG CTGCAGTCAG GCCATGGCGG CACTGCAGAA CT - #TGCCTCAG         720                                                                           - TGCTCCCCTG ATGAGATTAT GGCTTATGCC CAGAAAATAT TTAAGATTTT GG - #ATGAGGAG         780                                                                           - AGAGACAAGG TGCTCACGCA CATTGATCAC ATATTTATGG ATATCCTCAC TA - #CATGTGTG         840                                                                           - GAAACAATGT GTAATGAGTA CAAGGTCACT AGTGACGCTT GTATGATGAC CA - #TGTACGGG         900                                                                           - GGCATCTCTC TCTTAAGTGA GTTCTGTCGG GTGCTGTGCT GCTATGTCTT AG - #AGGAGACT         960                                                                           - AGTGTGATGC TGGCCAAGCG GCCTCTGATA ACCAAGCCTG AGGTTATCAG TG - #TAATGAAG        1020                                                                           - CGCCGCATTG AGGAGATCTG CATGAAGGTC TTTGCCCAGT ACATTCTGGG GG - #CCGATCCT        1080                                                                           - CTGAGAGTCT GCTCTCCTAG TGTGGATGAC CTACGGGCCA TCGCCGAGGA GT - #CAGATGAG        1140                                                                           - GAAGAGGCTA TTGTAGCCTA CACTTTGGCC ACCGCTGGTG TCAGCTCCTC TG - #ATTCTCTG        1200                                                                           - GTGTCACCCC CAGAGTCCCC TGTACCCGCG ACTATCCCTC TGTCCTCAGT AA - #TTGTGGCT        1260                                                                           - GAGAACAGTG ATCAGGAAGA AAGTGAGCAG AGTGATGAGG AAGAGGAGGA GG - #GTGCTCAG        1320                                                                           - GAGGAGCGGG AGGACACTGT GTCTGTCAAG TCTGAGCCAG TGTCTGAGAT AG - #AGGAAGTT        1380                                                                           - GCCCCAGAGG AAGAGGAGGA TGGTGCTGAG GAACCCACCG CCTCTGGAGG TA - #AGAGTACC        1440                                                                           #     1476         GAAG CAAGGCTGAC CAGTAA                                      - (2) INFORMATION FOR SEQ ID NO:51:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 1975 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:51:                                 - ATATAATCCT CCACCAAAAT AGAGAATATA TATATCATCA TTTCATGATG TA - #TACTACTG          60                                                                           - ACATAGTTTC AATGTGAACT TTTCACTTTC TTGCCGGTTA TGAAGAATAT TT - #TTTATTTT         120                                                                           - AATGGTCATT ACTAATCGTA TATTATAATT GAAAATGAAT TAGTTTAATA TG - #ACGCTCGT         180                                                                           - CATGGGATCC ATAAAAATTA CTGGTCAGCC TTGCTTCTAG TCACCATAGG GT - #GGGTACTC         240                                                                           - TTACCTCCAG AGGCGGTGGG TTCCTCAGCA CCATCCTCCT CTTCCTCTGG GG - #CAACTTCC         300                                                                           - TCTATCTCAG ACACTGGCTC AGACTTGACA GACACAGTGT CCTCCCGCTC CT - #CCTGAGCA         360                                                                           - CCCTCCTCCT CTTCCTCATC ACTCTGCTCA CTTTCTTCCT GATCACTGTT CT - #CAGCCACA         420                                                                           - ATTACTGAGG ACAGAGGGAT AGTCGCGGGT ACAGGGGACT CTGGGGGTGA CA - #CCAGAGAA         480                                                                           - TCAGAGGAGC TGACACCAGC GGTGGCCAAA GTGTAGGCTA CAATAGCCTC TT - #CCTCATCT         540                                                                           - GACTCCTCGG CGATGGCCCG TAGGTCATCC ACACTAGGAG AGCAGACTCT CA - #GAGGATCG         600                                                                           - GCCCCCAGAA TGTACTGGGC AAAGACCTTC ATGCAGATCT CCTCAATGCG GC - #GCTTCATT         660                                                                           - ACACTGATAA CCTCAGGCTT GGTTATCAGA GGCCGCTTGG CCAGCATCAC AC - #TAGTCTCC         720                                                                           - TCTAAGACAT AGCAGCACAG CACCCGACAG AACTCACTTA AGAGAGAGAT GC - #CCCCGTAC         780                                                                           - ATGGTCATCA TACAAGCGTC ACTAGTGACC TTGTACTCAT TACACATTGT TT - #CCACACAT         840                                                                           - GTAGTGAGGA TATCCATAAA TATGTGATCA ATGTGCGTGA GCACCTTGTC TC - #TCTCCTCA         900                                                                           - TCCAAAATCT TAAATATTTT CTGGGCATAA GCCATAATCT CATCAGGGGA GC - #ACTGAGGC         960                                                                           - AAGTTCTGCA GTGCCGCCAT GGCCTGACTG CAGCCATTGG TGGTCTTAGG GA - #AGGCTGAG        1020                                                                           - TTCTTGGTAA AGAACTCTAT ATTCCTGTAG CACATATACA TCATCTTTCT CC - #TAAGTTCA        1080                                                                           - TCCTTTTTAG CACGGGCCTT AGCCTGCAGT GCACCCCCCA ACTTGTTAGC GG - #CGCCCTTG        1140                                                                           - CTCACATCAT GCAGCTCCTT AATACAAGCC ATCCACATCT CCCGCTTATC CT - #CAGGTACA        1200                                                                           - ATGTAGTTCT CATACATGCT CTGCATAGTT AGCCCAATAC ACTTCATCTC CT - #CGAAAGGC        1260                                                                           - TCATGAACCT TATCTAAGAT ATCTAAGGCA TTCTGCAAAC ATCCTCCCAT CA - #TATTAAAG        1320                                                                           - GCGCCAGTGA ATTTCTCTTC CGTCTGGGTA TATTTTTTCA GCATGTGCTC CT - #TGATTCTA        1380                                                                           - TGCCGCACCA TGTCCACTCG AACCTTAATC TGTTTGACGA GTTCTGCCAG GA - #CATCTTTC        1440                                                                           - TCGGGGTTCT CGTTGCAATC CTCGGTCACT TGTTCAAAAG TTTTGAGGGA TT - #CTTCGGCC        1500                                                                           - AACTCTGGAA ACAGCGGGTC TCCCAGACTC AGCTGACTGT TAACCTCCTT CC - #TCAACATA        1560                                                                           - GTCTGCAGGA ACGTCGTGGC CTTGGTCACG GGTGTCTCGG GCCGTGGCAC CT - #TGGAGGAA        1620                                                                           - GGGCCCTCGT CAGGATTATC AGGGTCCATC TTTCTCTTGG CAGAGGACTC CA - #TTACGATA        1680                                                                           - CAAACTTAAC GGATATCGCG ATAATGAAAT AATTTATGAT TATTTCTCGC TT - #TCAATTTA        1740                                                                           - ACACAACCCT CAAGAACCTT TGTATTTATT TTCACTTTTT AAGTATAGAA TA - #AAGAGATC        1800                                                                           - CTGCTGTGGT AGATTCTGTG ACGCTAAGAA TAAGAATAAG AAGGAAGATG TA - #GAAGAGGG        1860                                                                           - AAGAGAAGGA TGTTACAATT ATAAGAACCT TAATGATCTG GATGAATCCG AA - #GCACGTGT        1920                                                                           - AGAATTTGGA CCATTATATA TGATAAATGA AGAAAAATCA GACATAAATA CA - #TTG             1975                                                                           - (2) INFORMATION FOR SEQ ID NO:52:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 3499 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52:                                 - AAGCTTGCGG CCGCTCATTA GACAAGCGAA TGAGGGACGA AAACGTGGAG GA - #GGTATTAA          60                                                                           - GTTTGGAGAA ATGGAGAGAG ACTGTTTAAT AGCGCATGGC GCAGCCAATA CT - #ATTACAGA         120                                                                           - AGTTTTGAAA GATTCGGAAG AAGATTATCA AGATGTGTAT GTTTGTGAAA AT - #TGTGGAGA         180                                                                           - CATAGCAGCA CAAATCAAGG GTATTAATAC ATGTCTTAGA TGTTCAAAAC TT - #AATCTCTC         240                                                                           - TCCTCTCTTA ACAAAAATTG ATACCACGCA CGTATCTAAA GTATTTCTTA CT - #CAAATGAA         300                                                                           - CGCCAGAGGC GTAAAAGTCA AATTAGATTT CGAACGAAGG CCTCCTTCGT TT - #TATAAACC         360                                                                           - ATTAGATAAA GTTGATCTCA AGCCGTCTTT TCTGGTGTAA TAAAAATTAA TT - #AATTACTC         420                                                                           - GAGATAAAAA TTACTGGTCA GCCTTGCTTC TAGTCACCAT AGGGTGGGTA CT - #CTTACCTC         480                                                                           - CAGAGGCGGT GGGTTCCTCA GCACCATCCT CCTCTTCCTC TGGGGCAACT TC - #CTCTATCT         540                                                                           - CAGACACTGG CTCAGACTTG ACAGACACAG TGTCCTCCCG CTCCTCCTGA GC - #ACCCTCCT         600                                                                           - CCTCTTCCTC ATCACTCTGC TCACTTTCTT CCTGATCACT GTTCTCAGCC AC - #AATTACTG         660                                                                           - AGGACAGAGG GATAGTCGCG GGTACAGGGG ACTCTGGGGG TGACACCAGA GA - #ATCAGAGG         720                                                                           - AGCTGACACC AGCGGTGGCC AAAGTGTAGG CTACAATAGC CTCTTCCTCA TC - #TGACTCCT         780                                                                           - CGGCGATGGC CCGTAGGTCA TCCACACTAG GAGAGCAGAC TCTCAGAGGA TC - #GGCCCCCA         840                                                                           - GAATGTACTG GGCAAAGACC TTCATGCAGA TCTCCTCAAT GCGGCGCTTC AT - #TACACTGA         900                                                                           - TAACCTCAGG CTTGGTTATC AGAGGCCGCT TGGCCAGCAT CACACTAGTC TC - #CTCTAAGA         960                                                                           - CATAGCAGCA CAGCACCCGA CAGAACTCAC TTAAGAGAGA GATGCCCCCG TA - #CATGGTCA        1020                                                                           - TCATACAAGC GTCACTAGTG ACCTTGTACT CATTACACAT TGTTTCCACA CA - #TGTAGTGA        1080                                                                           - GGATATCCAT AAATATGTGA TCAATGTGCG TGAGCACCTT GTCTCTCTCC TC - #ATCCAAAA        1140                                                                           - TCTTAAATAT TTTCTGGGCA TAAGCCATAA TCTCATCAGG GGAGCACTGA GG - #CAAGTTCT        1200                                                                           - GCAGTGCCGC CATGGCCTGA CTGCAGCCAT TGGTGGTCTT AGGGAAGGCT GA - #GTTCTTGG        1260                                                                           - TAAAGAACTC TATATTCCTG TAGCACATAT ACATCATCTT TCTCCTAAGT TC - #ATCCTTTT        1320                                                                           - TAGCACGGGC CTTAGCCTGC AGTGCACCCC CCAACTTGTT AGCGGCGCCC TT - #GCTCACAT        1380                                                                           - CATGCAGCTC CTTAATACAA GCCATCCACA TCTCCCGCTT ATCCTCAGGT AC - #AATGTAGT        1440                                                                           - TCTCATACAT GCTCTGCATA GTTAGCCCAA TACACTTCAT CTCCTCGAAA GG - #CTCATGAA        1500                                                                           - CCTTATCTAA GATATCTAAG GCATTCTGCA AACATCCTCC CATCATATTA AA - #GGCGCCAG        1560                                                                           - TGAATTTCTC TTCCGTCTGG GTATATTTTT TCAGCATGTG CTCCTTGATT CT - #ATGCCGCA        1620                                                                           - CCATGTCCAC TCGAACCTTA ATCTGTTTGA CGAGTTCTGC CAGGACATCT TT - #CTCGGGGT        1680                                                                           - TCTCGTTGCA ATCCTCGGTC ACTTGTTCAA AAGTTTTGAG GGATTCTTCG GC - #CAACTCTG        1740                                                                           - GAAACAGCGG GTCTCCCAGA CTCAGCTGAC TGTTAACCTC CTTCCTCAAC AT - #AGTCTGCA        1800                                                                           - GGAACGTCGT GGCCTTGGTC ACGGGTGTCT CGGGCCGTGG CACCTTGGAG GA - #AGGGCCCT        1860                                                                           - CGTCAGGATT ATCAGGGTCC ATCTTTCTCT TGGCAGAGGA CTCCATTACG AT - #ACAAACTT        1920                                                                           - AACGGATATC GCGATAATGA AATAATTTAT GATTATTTCT CGCTTTCAAT TT - #AACACAAC        1980                                                                           - CCTCAAGAAC CTTTGTATTT ATTTTCACTT TTTAAGTATA GAATAAAGAA GC - #TCTAATTA        2040                                                                           - ATTAAGCTAC AAATAGTTTC GTTTTCACCT TGTCTAATAA CTAATTAATT AA - #CCCCGATA        2100                                                                           - GCTGATTAGT TTTTGTTAAC AAAAATGTGG GAGAATCTAA TTAGTTTTTC TT - #TACACAAT        2160                                                                           - TGACGTACAT GAGTCTGAGT TCCTTGTTTT TGCTAATTAT TTCATCCAAT TT - #ATTATTCT        2220                                                                           - TGACGATATC GAGATCTTTT GTATAGGAGT CAGACTTGTA TTCAACATGC TT - #TTCTATAA        2280                                                                           - TCATCTTAGT TATTTCGGCA TCATCCAATA GTACATTTTC CAGATTAACA GA - #GTAGATAT        2340                                                                           - TAATGTCGTA TTTGAACAGA GCCTGTAACA TCTCAATGTC TTTATTATCT AT - #AGCCAATT        2400                                                                           - TAATGTCCGG AATGAAGAGA AGGGAATTAT TGGTGTTTGT CGACGTCATA TA - #GTCGAGCA        2460                                                                           - AGAGAATCAT CATATCCACG TGTCCATTTT TTATAGTGGT GTGAATACAA CT - #AAGGAGAA        2520                                                                           - TAGCCAGATC AAAAGTAGAT GGTATTTCTG AAAGAAAGTA TGATACAATA CT - #TACATCAT        2580                                                                           - TAAGCATGAC GGCATGATAA AATGAAGTTT TCCATCCAGT TTTCCCATAG AA - #CATCAGTC        2640                                                                           - TCCAATTTTT CTTAAACAGT TTCACCGTTT GCATGTTACC ACTATCAACC GC - #ATAATACA        2700                                                                           - ATGCGGTGTT TCCTTTGTCA TCAAATTGTG AATCATCCAT TCCACTGAAT AG - #CAAAATCT        2760                                                                           - TTACTATTTT GGTATCTTCT AATGTGGCTG CCTGATGTAA TGGAAATTCA TT - #CTCTAGAA        2820                                                                           - GATTTTTCAA TGCTCCAGCG TTCAACAACG TACATACTAG ACGCACGTTA TT - #ATCAGCTA        2880                                                                           - TTGCATAATA CAAGGCACTA TGTCCATGGA CATCCGCCTT AAATGTATCT TT - #ACTAGAGA        2940                                                                           - GAAAGCTTTT CAGCTGCTTA GACTTCCAAG TATTAATTCG TGACAGATCC AT - #GTCTGAAA        3000                                                                           - CGAGACGCTA ATTAGTGTAT ATTTTTTCAT TTTTTATAAT TTTGTCATAT TG - #CACCAGAA        3060                                                                           - TTATAATATC TCTAATAGAT CTAATTTAAT TTAATTTATA TAACTTATTT TT - #TGAATATA        3120                                                                           - CTTTTAATTA ACAAAAGAGT TAAGTTACTC ATATGGACGC CGTCCAGTCT GA - #ACATCAAT        3180                                                                           - CTTTTTAGCC AGAGATATCA TAGCCGCTCT TAGAGTTTCA GCGTGATTTT CC - #AACCTAAA        3240                                                                           - TAGAACTTCA TCGTTGCGTT TACAACACTT TTCTATTTGT TCAAACTTTG TT - #GTTACATT        3300                                                                           - AGTAATCTTT TTTTCCAAAT TAGTTAGCCG TTGTTTGAGA GTTTCCTCAT TG - #TCGTCTTG        3360                                                                           - CATCGGCTTT AACAATTGCT TCGCGTTTAG CCTCCTGGCT GTTCTTATCA GC - #CTTTGTAG        3420                                                                           - AAAAAAATTC AGTTGCTGGA ATTGCAAGAT CGTCATCTCC GGGGAAAAGA GT - #TCCGTCCA        3480                                                                           #                 349 - #9                                                     - (2) INFORMATION FOR SEQ ID NO:53:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 1386 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53:                                 - ATGGAGTCCT CTGCCAAGAG AAAGATGGAC CCTGATAATC CTGACGAGGG CC - #CTTCCTCC          60                                                                           - AAGGTGCCAC GGCCCGAGAC ACCCGTGACC AAGGCCACGA CGTTCCTGCA GA - #CTATGTTG         120                                                                           - AGGAAGGAGG TTAACAGTCA GCTGAGTCTG GGAGACCCGC TGTTTCCAGA GT - #TGGCCGAA         180                                                                           - GAATCCCTCA AAACTTTTGA ACAAGTGACC GAGGATTGCA ACGAGAACCC CG - #AGAAAGAT         240                                                                           - GTCCTGGCAG AACTCGTCAA ACAGATTAAG GTTCGAGTGG ACATGGTGCG GC - #ATAGAATC         300                                                                           - AAGGAGCACA TGCTGAAAAA ATATACCCAG ACGGAAGAGA AATTCACTGG CG - #CCTTTAAT         360                                                                           - ATGATGGGAG GATGTTTGCA GAATGCCTTA GATATCTTAG ATAAGGTTCA TG - #AGCCTTTC         420                                                                           - GAGGAGATGA AGTGTATTGG GCTAACTATG CAGAGCATGT ATGAGAACTA CA - #TTGTACCT         480                                                                           - GAGGATAAGC GGGAGATGTG GATGGCTTGT ATTAAGGAGC TGCATGATGT GA - #GCAAGGGC         540                                                                           - GCCGCTAACA AGTTGGGGGG TGCACTGCAG GCTAAGGCCC GTGCTAAAAA GG - #ATGAACTT         600                                                                           - AGGAGAAAGA TGATGTATAT GTGCTACAGG AATATAGAGT TCTTTACCAA GA - #ACTCAGCC         660                                                                           - TTCCCTAAGA CCACCAATGG CTGCAGTCAG GCCATGGCGG CACTGCAGAA CT - #TGCCTCAG         720                                                                           - TGCTCCCCTG ATGAGATTAT GGCTTATGCC CAGAAAATAT TTAAGATTTT GG - #ATGAGGAG         780                                                                           - AGAGACAAGG TGCTCACGCA CATTGATCAC ATATTTATGG ATATCCTCAC TA - #CATGTGTG         840                                                                           - GAAACAATGT GTAATGAGTA CAAGGTCACT AGTGTGATGC TGGCCAAGCG GC - #CTCTGATA         900                                                                           - ACCAAGCCTG AGGTTATCAG TGTAATGAAG CGCCGCATTG AGGAGATCTG CA - #TGAAGGTC         960                                                                           - TTTGCCCAGT ACATTCTGGG GGCCGATCCT CTGAGAGTCT GCTCTCCTAG TG - #TGGATGAC        1020                                                                           - CTACGGGCCA TCGCCGAGGA GTCAGATGAG GAAGAGGCTA TTGTAGCCTA CA - #CTTTGGCC        1080                                                                           - ACCGCTGGTG TCAGCTCCTC TGATTCTCTG GTGTCACCCC CAGAGTCCCC TG - #TACCCGCG        1140                                                                           - ACTATCCCTC TGTCCTCAGT AATTGTGGCT GAGAACAGTG ATCAGGAAGA AA - #GTGAGCAG        1200                                                                           - AGTGATGAGG AAGAGGAGGA GGGTGCTCAG GAGGAGCGGG AGGACACTGT GT - #CTGTCAAG        1260                                                                           - TCTGAGCCAG TGTCTGAGAT AGAGGAAGTT GCCCCAGAGG AAGAGGAGGA TG - #GTGCTGAG        1320                                                                           - GAACCCACCG CCTCTGGAGG TAAGAGTACC CACCCTATGG TGACTAGAAG CA - #AGGCTGAC        1380                                                                           #         1386                                                                 - (2) INFORMATION FOR SEQ ID NO:54:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 3409 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:54:                                 - AAGCTTGCGG CCGCTCATTA GACAAGCGAA TGAGGGACGA AAACGTGGAG GA - #GGTATTAA          60                                                                           - GTTTGGAGAA ATGGAGAGAG ACTGTTTAAT AGCGCATGGC GCAGCCAATA CT - #ATTACAGA         120                                                                           - AGTTTTGAAA GATTCGGAAG AAGATTATCA AGATGTGTAT GTTTGTGAAA AT - #TGTGGAGA         180                                                                           - CATAGCAGCA CAAATCAAGG GTATTAATAC ATGTCTTAGA TGTTCAAAAC TT - #AATCTCTC         240                                                                           - TCCTCTCTTA ACAAAAATTG ATACCACGCA CGTATCTAAA GTATTTCTTA CT - #CAAATGAA         300                                                                           - CGCCAGAGGC GTAAAAGTCA AATTAGATTT CGAACGAAGG CCTCCTTCGT TT - #TATAAACC         360                                                                           - ATTAGATAAA GTTGATCTCA AGCCGTCTTT TCTGGTGTAA TAAAAATTAA TT - #AATTACTC         420                                                                           - GAGATAAAAA TTACTGGTCA GCCTTGCTTC TAGTCACCAT AGGGTGGGTA CT - #CTTACCTC         480                                                                           - CAGAGGCGGT GGGTTCCTCA GCACCATCCT CCTCTTCCTC TGGGGCAACT TC - #CTCTATCT         540                                                                           - CAGACACTGG CTCAGACTTG ACAGACACAG TGTCCTCCCG CTCCTCCTGA GC - #ACCCTCCT         600                                                                           - CCTCTTCCTC ATCACTCTGC TCACTTTCTT CCTGATCACT GTTCTCAGCC AC - #AATTACTG         660                                                                           - AGGACAGAGG GATAGTCGCG GGTACAGGGG ACTCTGGGGG TGACACCAGA GA - #ATCAGAGG         720                                                                           - AGCTGACACC AGCGGTGGCC AAAGTGTAGG CTACAATAGC CTCTTCCTCA TC - #TGACTCCT         780                                                                           - CGGCGATGGC CCGTAGGTCA TCCACACTAG GAGAGCAGAC TCTCAGAGGA TC - #GGCCCCCA         840                                                                           - GAATGTACTG GGCAAAGACC TTCATGCAGA TCTCCTCAAT GCGGCGCTTC AT - #TACACTGA         900                                                                           - TAACCTCAGG CTTGGTTATC AGAGGCCGCT TGGCCAGCAT CACACTAGTG AC - #CTTGTACT         960                                                                           - CATTACACAT TGTTTCCACA CATGTAGTGA GGATATCCAT AAATATGTGA TC - #AATGTGCG        1020                                                                           - TGAGCACCTT GTCTCTCTCC TCATCCAAAA TCTTAAATAT TTTCTGGGCA TA - #AGCCATAA        1080                                                                           - TCTCATCAGG GGAGCACTGA GGCAAGTTCT GCAGTGCCGC CATGGCCTGA CT - #GCAGCCAT        1140                                                                           - TGGTGGTCTT AGGGAAGGCT GAGTTCTTGG TAAAGAACTC TATATTCCTG TA - #GCACATAT        1200                                                                           - ACATCATCTT TCTCCTAAGT TCATCCTTTT TAGCACGGGC CTTAGCCTGC AG - #TGCACCCC        1260                                                                           - CCAACTTGTT AGCGGCGCCC TTGCTCACAT CATGCAGCTC CTTAATACAA GC - #CATCCACA        1320                                                                           - TCTCCCGCTT ATCCTCAGGT ACAATGTAGT TCTCATACAT GCTCTGCATA GT - #TAGCCCAA        1380                                                                           - TACACTTCAT CTCCTCGAAA GGCTCATGAA CCTTATCTAA GATATCTAAG GC - #ATTCTGCA        1440                                                                           - AACATCCTCC CATCATATTA AAGGCGCCAG TGAATTTCTC TTCCGTCTGG GT - #ATATTTTT        1500                                                                           - TCAGCATGTG CTCCTTGATT CTATGCCGCA CCATGTCCAC TCGAACCTTA AT - #CTGTTTGA        1560                                                                           - CGAGTTCTGC CAGGACATCT TTCTCGGGGT TCTCGTTGCA ATCCTCGGTC AC - #TTGTTCAA        1620                                                                           - AAGTTTTGAG GGATTCTTCG GCCAACTCTG GAAACAGCGG GTCTCCCAGA CT - #CAGCTGAC        1680                                                                           - TGTTAACCTC CTTCCTCAAC ATAGTCTGCA GGAACGTCGT GGCCTTGGTC AC - #GGGTGTCT        1740                                                                           - CGGGCCGTGG CACCTTGGAG GAAGGGCCCT CGTCAGGATT ATCAGGGTCC AT - #CTTTCTCT        1800                                                                           - TGGCAGAGGA CTCCATTACG ATACAAACTT AACGGATATC GCGATAATGA AA - #TAATTTAT        1860                                                                           - GATTATTTCT CGCTTTCAAT TTAACACAAC CCTCAAGAAC CTTTGTATTT AT - #TTTCACTT        1920                                                                           - TTTAAGTATA GAATAAAGAA GCTCTAATTA ATTAAGCTAC AAATAGTTTC GT - #TTTCACCT        1980                                                                           - TGTCTAATAA CTAATTAATT AACCCCGATA GCTGATTAGT TTTTGTTAAC AA - #AAATGTGG        2040                                                                           - GAGAATCTAA TTAGTTTTTC TTTACACAAT TGACGTACAT GAGTCTGAGT TC - #CTTGTTTT        2100                                                                           - TGCTAATTAT TTCATCCAAT TTATTATTCT TGACGATATC GAGATCTTTT GT - #ATAGGAGT        2160                                                                           - CAGACTTGTA TTCAACATGC TTTTCTATAA TCATCTTAGT TATTTCGGCA TC - #ATCCAATA        2220                                                                           - GTACATTTTC CAGATTAACA GAGTAGATAT TAATGTCGTA TTTGAACAGA GC - #CTGTAACA        2280                                                                           - TCTCAATGTC TTTATTATCT ATAGCCAATT TAATGTCCGG AATGAAGAGA AG - #GGAATTAT        2340                                                                           - TGGTGTTTGT CGACGTCATA TAGTCGAGCA AGAGAATCAT CATATCCACG TG - #TCCATTTT        2400                                                                           - TTATAGTGGT GTGAATACAA CTAAGGAGAA TAGCCAGATC AAAAGTAGAT GG - #TATTTCTG        2460                                                                           - AAAGAAAGTA TGATACAATA CTTACATCAT TAAGCATGAC GGCATGATAA AA - #TGAAGTTT        2520                                                                           - TCCATCCAGT TTTCCCATAG AACATCAGTC TCCAATTTTT CTTAAACAGT TT - #CACCGTTT        2580                                                                           - GCATGTTACC ACTATCAACC GCATAATACA ATGCGGTGTT TCCTTTGTCA TC - #AAATTGTG        2640                                                                           - AATCATCCAT TCCACTGAAT AGCAAAATCT TTACTATTTT GGTATCTTCT AA - #TGTGGCTG        2700                                                                           - CCTGATGTAA TGGAAATTCA TTCTCTAGAA GATTTTTCAA TGCTCCAGCG TT - #CAACAACG        2760                                                                           - TACATACTAG ACGCACGTTA TTATCAGCTA TTGCATAATA CAAGGCACTA TG - #TCCATGGA        2820                                                                           - CATCCGCCTT AAATGTATCT TTACTAGAGA GAAAGCTTTT CAGCTGCTTA GA - #CTTCCAAG        2880                                                                           - TATTAATTCG TGACAGATCC ATGTCTGAAA CGAGACGCTA ATTAGTGTAT AT - #TTTTTCAT        2940                                                                           - TTTTTATAAT TTTGTCATAT TGCACCAGAA TTAATAATAT CTCTAATAGA TC - #TAATTTAA        3000                                                                           - TTTAATTTAT ATAACTTATT TTTTGAATAT ACTTTTAATT AACAAAAGAG TT - #AAGTTACT        3060                                                                           - CATATGGACG CCGTCCAGTC TGAACATCAA TCTTTTTAGC CAGAGATATC AT - #AGCCGCTC        3120                                                                           - TTAGAGTTTC AGCGTGATTT TCCAACCTAA ATAGAACTTC ATCGTTGCGT TT - #ACAACACT        3180                                                                           - TTTCTATTTG TTCAAACTTT GTTGTTACAT TAGTAATCTT TTTTTCCAAA TT - #AGTTAGCC        3240                                                                           - GTTGTTTGAG AGTTTCCTCA TTGTCGTCTT CATCGGCTTT AACAATTGCT TC - #GCGTTTAG        3300                                                                           - CCTCCTGGCT GTTCTTATCA GCCTTTGTAG AAAAAAATTC AGTTGCTGGA AT - #TGCAAGAT        3360                                                                           #             3409AAAGA GTTCCGTCCA TTTAAAGCCG CGGGAATTC                        - (2) INFORMATION FOR SEQ ID NO:55:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 1221 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:55:                                 - ATGAAACAGA TTAAGGTTCG AGTGGACATG GTGCGGCATA GAATCAAGGA GC - #ACATGCTG          60                                                                           - AAAAAATATA CCCAGACGGA AGAGAAATTC ACTGGCGCCT TTAATATGAT GG - #GAGGATGT         120                                                                           - TTGCAGAATG CCTTAGATAT CTTAGATAAG GTTCATGAGC CTTTCGAGGA GA - #TGAAGTGT         180                                                                           - ATTGGGCTAA CTATGCAGAG CATGTATGAG AACTACATTG TACCTGAGGA TA - #AGCGGGAG         240                                                                           - ATGTGGATGG CTTGTATTAA GGAGCTGCAT GATGTGAGCA AGGGCGCCGC TA - #ACAAGTTG         300                                                                           - GGGGGTGCAC TGCAGGCTAA GGCCCGTGCT AAAAAGGATG AACTTAGGAG AA - #AGATGATG         360                                                                           - TATATGTGCT ACAGGAATAT AGAGTTCTTT ACCAAGAACT CAGCCTTCCC TA - #AGACCACC         420                                                                           - AATGGCTGCA GTCAGGCCAT GGCGGCACTG CAGAACTTGC CTCAGTGCTC CC - #CTGATGAG         480                                                                           - ATTATGGCTT ATGCCCAGAA AATATTTAAG ATTTTGGATG AGGAGAGAGA CA - #AGGTGCTC         540                                                                           - ACGCACATTG ATCACATATT TATGGATATC CTCACTACAT GTGTGGAAAC AA - #TGTGTAAT         600                                                                           - GAGTACAAGG TCACTAGTGA CGCTTGTATG ATGACCATGT ACGGGGGCAT CT - #CTCTCTTA         660                                                                           - AGTGAGTTCT GTCGGGTGCT GTGCTGCTAT GTCTTAGAGG AGACTAGTGT GA - #TGCTGGCC         720                                                                           - AAGCGGCCTC TGATAACCAA GCCTGAGGTT ATCAGTGTAA TGAAGCGCCG CA - #TTGAGGAG         780                                                                           - ATCTGCATGA AGGTCTTTGC CCAGTACATT CTGGGGGCCG ATCCTCTGAG AG - #TCTGCTCT         840                                                                           - CCTAGTGTGG ATGACCTACG GGCCATCGCC GAGGAGTCAG ATGAGGAAGA GG - #CTATTGTA         900                                                                           - GCCTACACTT TGGCCACCGC TGGTGTCAGC TCCTCTGATT CTCTGGTGTC AC - #CCCCAGAG         960                                                                           - TCCCCTGTAC CCGCGACTAT CCCTCTGTCC TCAGTAATTG TGGCTGAGAA CA - #GTGATCAG        1020                                                                           - GAAGAAAGTG AGCAGAGTGA TGAGGAAGAG GAGGAGGGTG CTCAGGAGGA GC - #GGGAGGAC        1080                                                                           - ACTGTGTCTG TCAAGTCTGA GCCAGTGTCT GAGATAGAGG AAGTTGCCCC AG - #AGGAAGAG        1140                                                                           - GAGGATGGTG CTGAGGAACC CACCGCCTCT GGAGGTAAGA GTACCCACCC TA - #TGGTGACT        1200                                                                           #                1221TA A                                                      - (2) INFORMATION FOR SEQ ID NO:56:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 2577 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:56:                                 - CTGCAGGTCG ACGGATCTGA GAATGGATGA TTCTCCAGCC GAAACATATT CT - #ACCATGGC          60                                                                           - TCCGTTTAAT TTGTTGATGA AGATGGATTC ATCCTTAAAT GTTTTCTCTG TA - #ATAGTTTC         120                                                                           - CACCGAAAGA CTATGCAAAG AATTTGGAAT GCGTTCCTTG TGCTTAATGT TT - #CCATAGAC         180                                                                           - GGCTTCTAGA AGTTGATACA ACATAGGACT AGCCGCGGTA ACTTTTATTT TT - #AGAAAGTA         240                                                                           - TCCATCGCTT CTATCTTGTT TAGATTTATT TTTATAAAGT TTAGTCTCTC CT - #TCCAACAT         300                                                                           - AATAAAAGTG GAAGTCATTT GACTAGATAA ACTATCAGTA AGTTTTATAG AG - #ATAGACGA         360                                                                           - ACAATTAGCG TATTGAGAAG CATTTAGTGT AACGTATTCG ATACATTTTG CA - #TTAGATTT         420                                                                           - ACTAATCGAT TTTGCATACT CTATAACACC CGCACAAGTC TGTAGAGAAT CG - #CTAGATGC         480                                                                           - AGTAGGTCTT GGTGAAGTTT CAACTCTCTT CTTGATTACC TTACTCATGA TT - #AAACCTAA         540                                                                           - ATAATTGTAC TTTGTAATAT AATGATATAT ATTTTCACTT TATCTCATTT GA - #GAATAAAA         600                                                                           - AGATCACAAA AATTAACTAA TCAGGATCCT TCTTTATTCT ATACTTAAAA AG - #TGAAAATA         660                                                                           - AATACAAAGG TTCTTGAGGG TTGTGTTAAA TTGAAAGCGA GAAATAATCA TA - #AATTATTT         720                                                                           - CATTATCGCG ATATCCGTTA AGTTTGTATC GTAATGAAAC AGATTAAGGT TC - #GAGTGGAC         780                                                                           - ATGGTGCGGC ATAGAATCAA GGAGCACATG CTGAAAAAAT ATACCCAGAC GG - #AAGAGAAA         840                                                                           - TTCACTGGCG CCTTTAATAT GATGGGAGGA TGTTTGCAGA ATGCCTTAGA TA - #TCTTAGAT         900                                                                           - AAGGTTCATG AGCCTTTCGA GGAGATGAAG TGTATTGGGC TAACTATGCA GA - #GCATGTAT         960                                                                           - GAGAACTACA TTGTACCTGA GGATAAGCGG GAGATGTGGA TGGCTTGTAT TA - #AGGAGCTG        1020                                                                           - CATGATGTGA GCAAGGGCGC CGCTAACAAG TTGGGGGGTG CACTGCAGGC TA - #AGGCCCGT        1080                                                                           - GCTAAAAAGG ATGAACTTAG GAGAAAGATG ATGTATATGT GCTACAGGAA TA - #TAGAGTTC        1140                                                                           - TTTACCAAGA ACTCAGCCTT CCCTAAGACC ACCAATGGCT GCAGTCAGGC CA - #TGGCGGCA        1200                                                                           - CTGCAGAACT TGCCTCAGTG CTCCCCTGAT GAGATTATGG CTTATGCCCA GA - #AAATATTT        1260                                                                           - AAGATTTTGG ATGAGGAGAG AGACAAGGTG CTCACGCACA TTGATCACAT AT - #TTATGGAT        1320                                                                           - ATCCTCACTA CATGTGTGGA AACAATGTGT AATGAGTACA AGGTCACTAG TG - #ACGCTTGT        1380                                                                           - ATGATGACCA TGTACGGGGG CATCTCTCTC TTAAGTGAGT TCTGTCGGGT GC - #TGTGCTGC        1440                                                                           - TATGTCTTAG AGGAGACTAG TGTGATGCTG GCCAAGCGGC CTCTGATAAC CA - #AGCCTGAG        1500                                                                           - GTTATCAGTG TAATGAAGCG CCGCATTGAG GAGATCTGCA TGAAGGTCTT TG - #CCCAGTAC        1560                                                                           - ATTCTGGGGG CCGATCCTCT GAGAGTCTGC TCTCCTAGTG TGGATGACCT AC - #GGGCCATC        1620                                                                           - GCCGAGGAGT CAGATGAGGA AGAGGCTATT GTAGCCTACA CTTTGGCCAC CG - #CTGGTGTC        1680                                                                           - AGCTCCTCTG ATTCTCTGGT GTCACCCCCA GAGTCCCCTG TACCCGCGAC TA - #TCCCTCTG        1740                                                                           - TCCTCAGTAA TTGTGGCTGA GAACAGTGAT CAGGAAGAAA GTGAGCAGAG TG - #ATGAGGAA        1800                                                                           - GAGGAGGAGG GTGCTCAGGA GGAGCGGGAG GACACTGTGT CTGTCAAGTC TG - #AGCCAGTG        1860                                                                           - TCTGAGATAG AGGAAGTTGC CCCAGAGGAA GAGGAGGATG GTGCTGAGGA AC - #CCACCGCC        1920                                                                           - TCTGGAGGTA AGAGTACCCA CCCTATGGTG ACTAGAAGCA AGGCTGACCA GT - #AATTTTTA        1980                                                                           - TCTCGAGCCC GGGAGATCTT AGCTAACTGA TTTTTCTGGG AAAAAAATTA TT - #TAACTTTT        2040                                                                           - CATTAATAGG GATTTGACGT ATGTAGCGTA CAAAATTATC GTTCCTGGTA TA - #TAGATAAA        2100                                                                           - GAGTCCTATA TATTTGAAAA TCGTTACGGC TCGATTAAAC TTTAATGATT GC - #ATAGTGAA        2160                                                                           - TATATCATTA GGATTTAACT CCTTGACTAT CATGGCGGCG CCAGAAATTA CC - #ATCAAAAG        2220                                                                           - CATTAATACA GTTATGCCGA TCGCAGTTAG AACGGTTATA GCATCCACCA TT - #TATATCTA        2280                                                                           - AAAATTAGAT CAAAGAATAT GTGACAAAGT CCTAGTTGTA TACTGAGAAT TG - #ACGAAACA        2340                                                                           - ATGTTTCTTA CATATTTTTT TCTTATTAGT AACTGACTTA ATAGTAGGAA CT - #GGAAAGCT        2400                                                                           - AGACTTGATT ATTCTATAAG TATAGATACC CTTCCAGATA ATGTTCTCTT TG - #ATAAAAGT        2460                                                                           - TCCAGAAAAT GTAGAATTTT TTAAAAAGTT ATCTTTTGCT ATTACCAAGA TT - #GTGTTTAG        2520                                                                           - ACGCTTATTA TTAATATGAG TAATGAAATC CACACCGCCT CTAGATATGG GG - #AATTC           2577                                                                           - (2) INFORMATION FOR SEQ ID NO:57:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 3460 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57:                                 - GAATTGCGGC CGCTGAATGT TAAATGTTAT ACTTTGGATG AAGCTATAAA TA - #TGCATTGG          60                                                                           - AAAAATAATC CATTTAAAGA AAGGATTCAA ATACTACAAA ACCTAAGCGA TA - #ATATGTTA         120                                                                           - ACTAAGCTTA TTCTTAACGA CGCTTTAAAT ATACACAAAT AAACATAATT TT - #TGTATAAC         180                                                                           - CTAACAAATA ACTAAAACAT AAAAATAATA AAAGGAAATG TAATATCGTA AT - #TATTTTAC         240                                                                           - TCAGGAATGG GGTTAAATAT TTATATCACG TGTATATCTA TACTGTTATC GT - #ATACTCTT         300                                                                           - TACAATTACT ATTACGAATA TGCAAGAGAT AATAAGATTA CGTATTTAAG AG - #AATCTTGT         360                                                                           - CATGATAATT GGGTACGACA TAGTGATAAA TGCTATTTCG CATCGTTACA TA - #AAGTCAGT         420                                                                           - TGGAAAGATG GATTTGACAG ATGTAACTTA ATAGGTGCAA AAATGTTAAA TA - #ACAGCATT         480                                                                           - CTATCGGAAG ATAGGATACC AGTTATATTA TACAAAAATC ACTGGTTGGA TA - #AAACAGAT         540                                                                           - TCTGCAATAT TCGTAAAAGA TGAAGATTAC TGCGAATTTG TAAACTATGA CA - #ATAAAAAG         600                                                                           - CCATTTATCT CAACGACATC GTGTAATTCT TCCATGTTTT ATGTATGTGT TT - #CAGATATT         660                                                                           - ATGAGATTAC TATAAACTTT TTGTATACTT ATATTCCGTA AACTATATTA AT - #CATGAAGA         720                                                                           - AAATGAAAAA GTATAGAAGC TGTTCACGAG CGGTTGTTGA AAACAACAAA AT - #TATACATT         780                                                                           - CAAGATGGCT TACATATACG TCTGTGAGGC TATCATGGAT AATGACAATG CA - #TCTCTAAA         840                                                                           - TAGGTTTTTG GACAATGGAT TCGACCCTAA CACGGAATAT GGTACTCTAC AA - #TCTCCTCT         900                                                                           - TGAAATGGCT GTAATGTTCA AGAATACCGA GGCTATAAAA ATCTTGATGA GG - #TATGGAGC         960                                                                           - TAAACCTGTA GTTACTGAAT GCACAACTTC TTGTCTGCAT GATGCGGTGT TG - #AGAGACGA        1020                                                                           - CTACAAAATA GTGAAAGATC TGTTGAAGAA TAACTATGTA AACAATGTTC TT - #TACAGCGG        1080                                                                           - AGGCTTTACT CCTTTGTGTT TGGCAGCTTA CCTTAACAAA GTTAATTTGG TT - #AAACTTCT        1140                                                                           - ATTGGCTCAT TCGGCGGATG TAGATATTTC AAACACGGAT CGGTTAACTC CT - #CTACATAT        1200                                                                           - AGCCGTATCA AATAAAAATT TAACAATGGT TAAACTTCTA TTGAACAAAG GT - #GCTGATAC        1260                                                                           - TGACTTGCTG GATAACATGG GACGTACTCC TTTAATGATC GCTGTACAAT CT - #GGAAATAT        1320                                                                           - TGAAATATGT AGCACACTAC TTAAAAAAAA TAAAATGTCC AGAACTGGGA AA - #AATTGATC        1380                                                                           - TTGCCAGCTG TAATTCATGG TAGAAAAGAA GTGCTCAGGC TACTTTTCAA CA - #AAGGAGCA        1440                                                                           - GATGTAAACT ACATCTTTGA AAGAAATGGA AAATCATATA CTGTTTTGGA AT - #TGATTAAA        1500                                                                           - GAAAGTTACT CTGAGACACA AAAGAGGTAG CTGAAGTGGT ACTCTCAAAG GT - #ACGTGACT        1560                                                                           - AATTAGCTAT AAAAAGGATC CGGGTTAATT AATTAGTCAT CAGGCAGGGC GA - #GAACGAGA        1620                                                                           - CTATCTGCTC GTTAATTAAT TAGAGCTTCT TTATTCTATA CTTAAAAAGT GA - #AAATAAAT        1680                                                                           - ACAAAGGTTC TTGAGGGTTG TGTTAAATTG AAAGCGAGAA ATAATCATAA AT - #TATTTCAT        1740                                                                           - TATCGCGATA TCCGTTAAGT TTGTATCGTA ATGAAACAGA TTAAGGTTCG AG - #TGGACATG        1800                                                                           - GTGCGGCATA GAATCAAGGA GCACATGCTG AAAAAATATA CCCAGACGGA AG - #AGAAATTC        1860                                                                           - ACTGGCGCCT TTAATATGAT GGGAGGATGT TTGCAGAATG CCTTAGATAT CT - #TAGATAAG        1920                                                                           - GTTCATGAGC CTTTCGAGGA GATGAAGTGT ATTGGGCTAA CTATGCAGAG CA - #TGTATGAG        1980                                                                           - AACTACATTG TACCTGAGGA TAAGCGGGAG ATGTGGATGG CTTGTATTAA GG - #AGCTGCAT        2040                                                                           - GATGTGAGCA AGGGCGCCGC TAACAAGTTG GGGGGTGCAC TGCAGGCTAA GG - #CCCGTGCT        2100                                                                           - AAAAAGGATG AACTTAGGAG AAAGATGATG TATATGTGCT ACAGGAATAT AG - #AGTTCTTT        2160                                                                           - ACCAAGAACT CAGCCTTCCC TAAGACCACC AATGGCTGCA GTCAGGCCAT GG - #CGGCACTG        2220                                                                           - CAGAACTTGC CTCAGTGCTC CCCTGATGAG ATTATGGCTT ATGCCCAGAA AA - #TATTTAAG        2280                                                                           - ATTTTGGATG AGGAGAGAGA CAAGGTGCTC ACGCACATTG ATCACATATT TA - #TGGATATC        2340                                                                           - CTCACTACAT GTGTGGAAAC AATGTGTAAT GAGTACAAGG TCACTAGTGA CG - #CTTGTATG        2400                                                                           - ATGACCATGT ACGGGGGCAT CTCTCTCTTA AGTGAGTTCT GTCGGGTGCT GT - #GCTGCTAT        2460                                                                           - GTCTTAGAGG AGACTAGTGT GATGCTGGCC AAGCGGCCTC TGATAACCAA GC - #CTGAGGTT        2520                                                                           - ATCAGTGTAA TGAAGCGCCG CATTGAGGAG ATCTGCATGA AGGTCTTTGC CC - #AGTACATT        2580                                                                           - CTGGGGGCCG ATCCTCTGAG AGTCTGCTCT CCTAGTGTGG ATGACCTACG GG - #CCATCGCC        2640                                                                           - GAGGAGTCAG ATGAGGAAGA GGCTATTGTA GCCTACACTT TGGCCACCGC TG - #GTGTCAGC        2700                                                                           - TCCTCTGATT CTCTGGTGTC ACCCCCAGAG TCCCCTGTAC CCGCGACTAT CC - #CTCTGTCC        2760                                                                           - TCAGTAATTG TGGCTGAGAA CAGTGATCAG GAAGAAAGTG AGCAGAGTGA TG - #AGGAAGAG        2820                                                                           - GAGGAGGGTG CTCAGGAGGA GCGGGAGGAC ACTGTGTCTG TCAAGTCTGA GC - #CAGTGTCT        2880                                                                           - GAGATAGAGG AAGTTGCCCC AGAGGAAGAG GAGGATGGTG CTGAGGAACC CA - #CCGCCTCT        2940                                                                           - GGAGGTAAGA GTACCCACCC TATGGTGACT AGAAGCAAGG CTGACCAGTA AT - #TTTTATCT        3000                                                                           - CGAGTCTAGA ATCGATCCCG GGTTTTTATG ACTAGTTAAT CACGGCCGCT TA - #TAAAGATC        3060                                                                           - TAAAATGCAT AATTTCTAAA TAATGAAAAA AAAGTACATC ATGAGCAACG CG - #TTAGTATA        3120                                                                           - TTTTACAATG GAGATTAACG CTCTATACCG TTCTATGTTT ATTGATTCAG AT - #GATGTTTT        3180                                                                           - AGAAAAGAAA GTTATTGAAT ATGAAAACTT TAATGAAGAT GAAGATGACG AC - #GATGATTA        3240                                                                           - TTGTTGTAAA TCTGTTTTAG ATGAAGAAGA TGACGCGCTA AAGTATACTA TG - #GTTACAAA        3300                                                                           - GTATAAGTCT ATACTACTAA TGGCGACTTG TGCAAGAAGG TATAGTATAG TG - #AAAATGTT        3360                                                                           - GTTAGATTAT GATTATGAAA AACCAAATAA ATCAGATCCA TATCTAAAGG TA - #TCTCCTTT        3420                                                                           #  3460            ATTC CTAGTTTAGA ATACCTGCAG                                  - (2) INFORMATION FOR SEQ ID NO:58:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 1383 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58:                                 - ATGACGACGT TCCTGCAGAC TATGTTGAGG AAGGAGGTTA ACAGTCAGCT GA - #GTCTGGGA          60                                                                           - GACCCGCTGT TTCCAGAGTT GGCCGAAGAA TCCCTCAAAA CTTTTGAACA AG - #TGACCGAG         120                                                                           - GATTGCAACG AGAACCCCGA GAAAGATGTC CTGGCAGAAC TCGTCAAACA GA - #TTAAGGTT         180                                                                           - CGAGTGGACA TGGTGCGGCA TAGAATCAAG GAGCACATGC TGAAAAAATA TA - #CCCAGACG         240                                                                           - GAAGAGAAAT TCACTGGCGC CTTTAATATG ATGGGAGGAT GTTTGCAGAA TG - #CCTTAGAT         300                                                                           - ATCTTAGATA AGGTTCATGA GCCTTTCGAG GAGATGAAGT GTATTGGGCT AA - #CTATGCAG         360                                                                           - AGCATGTATG AGAACTACAT TGTACCTGAG GATAAGCGGG AGATGTGGAT GG - #CTTGTATT         420                                                                           - AAGGAGCTGC ATGATGTGAG CAAGGGCGCC GCTAACAAGT TGGGGGGTGC AC - #TGCAGGCT         480                                                                           - AAGGCCCGTG CTAAAAAGGA TGAACTTAGG AGAAAGATGA TGTATATGTG CT - #ACAGGAAT         540                                                                           - ATAGAGTTCT TTACCAAGAA CTCAGCCTTC CCTAAGACCA CCAATGGCTG CA - #GTCAGGCC         600                                                                           - ATGGCGGCAC TGCAGAACTT GCCTCAGTGC TCCCCTGATG AGATTATGGC TT - #ATGCCCAG         660                                                                           - AAAATATTTA AGATTTTGGA TGAGGAGAGA GACAAGGTGC TCACGCACAT TG - #ATCACATA         720                                                                           - TTTATGGATA TCCTCACTAC ATGTGTGGAA ACAATGTGTA ATGAGTACAA GG - #TCACTAGT         780                                                                           - GACGCTTGTA TGATGACCAT GTACGGGGGC ATCTCTCTCT TAAGTGAGTT CT - #GTCGGGTG         840                                                                           - CTGTGCTGCT ATGTCTTAGA GGAGACTAGT GTGATGCTGG CCAAGCGGCC TC - #TGATAACC         900                                                                           - AAGCCTGAGG TTATCAGTGT AATGAAGCGC CGCATTGAGG AGATCTGCAT GA - #AGGTCTTT         960                                                                           - GCCCAGTACA TTCTGGGGGC CGATCCTCTG AGAGTCTGCT CTCCTAGTGT GG - #ATGACCTA        1020                                                                           - CGGGCCATCG CCGAGGAGTC AGATGAGGAA GAGGCTATTG TAGCCTACAC TT - #TGGCCACC        1080                                                                           - GCTGGTGTCA GCTCCTCTGA TTCTCTGGTG TCACCCCCAG AGTCCCCTGT AC - #CCGCGACT        1140                                                                           - ATCCCTCTGT CCTCAGTAAT TGTGGCTGAG AACAGTGATC AGGAAGAAAG TG - #AGCAGAGT        1200                                                                           - GATGAGGAAG AGGAGGAGGG TGCTCAGGAG GAGCGGGAGG ACACTGTGTC TG - #TCAAGTCT        1260                                                                           - GAGCCAGTGT CTGAGATAGA GGAAGTTGCC CCAGAGGAAG AGGAGGATGG TG - #CTGAGGAA        1320                                                                           - CCCACCGCCT CTGGAGGTAA GAGTACCCAC CCTATGGTGA CTAGAAGCAA GG - #CTGACCAG        1380                                                                           #           1383                                                               - (2) INFORMATION FOR SEQ ID NO:59:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 2739 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:59:                                 - CTGCAGGTCG ACGGATCTGA GAATGGATGA TTCTCCAGCC GAAACATATT CT - #ACCATGGC          60                                                                           - TCCGTTTAAT TTGTTGATGA AGATGGATTC ATCCTTAAAT GTTTTCTCTG TA - #ATAGTTTC         120                                                                           - CACCGAAAGA CTATGCAAAG AATTTGGAAT GCGTTCCTTG TGCTTAATGT TT - #CCATAGAC         180                                                                           - GGCTTCTAGA AGTTGATACA ACATAGGACT AGCCGCGGTA ACTTTTATTT TT - #AGAAAGTA         240                                                                           - TCCATCGCTT CTATCTTGTT TAGATTTATT TTTATAAAGT TTAGTCTCTC CT - #TCCAACAT         300                                                                           - AATAAAAGTG GAAGTCATTT GACTAGATAA ACTATCAGTA AGTTTTATAG AG - #ATAGACGA         360                                                                           - ACAATTAGCG TATTGAGAAG CATTTAGTGT AACGTATTCG ATACATTTTG CA - #TTAGATTT         420                                                                           - ACTAATCGAT TTTGCATACT CTATAACACC CGCACAAGTC TGTAGAGAAT CG - #CTAGATGC         480                                                                           - AGTAGGTCTT GGTGAAGTTT CAACTCTCTT CTTGATTACC TTACTCATGA TT - #AAACCTAA         540                                                                           - ATAATTGTAC TTTGTAATAT AATGATATAT ATTTTCACTT TATCTCATTT GA - #GAATAAAA         600                                                                           - AGATCACAAA AATTAACTAA TCAGGATCCT TCTTTATTCT ATACTTAAAA AG - #TGAAAATA         660                                                                           - AATACAAAGG TTCTTGAGGG TTGTGTTAAA TTGAAAGCGA GAAATAATCA TA - #AATTATTT         720                                                                           - CATTATCGCG ATATCCGTTA AGTTTGTATC GTAATGACGA CGTTCCTGCA GA - #CTATGTTG         780                                                                           - AGGAAGGAGG TTAACAGTCA GCTGAGTCTG GGAGACCCGC TGTTTCCAGA GT - #TGGCCGAA         840                                                                           - GAATCCCTCA AAACTTTTGA ACAAGTGACC GAGGATTGCA ACGAGAACCC CG - #AGAAAGAT         900                                                                           - GTCCTGGCAG AACTCGTCAA ACAGATTAAG GTTCGAGTGG ACATGGTGCG GC - #ATAGAATC         960                                                                           - AAGGAGCACA TGCTGAAAAA ATATACCCAG ACGGAAGAGA AATTCACTGG CG - #CCTTTAAT        1020                                                                           - ATGATGGGAG GATGTTTGCA GAATGCCTTA GATATCTTAG ATAAGGTTCA TG - #AGCCTTTC        1080                                                                           - GAGGAGATGA AGTGTATTGG GCTAACTATG CAGAGCATGT ATGAGAACTA CA - #TTGTACCT        1140                                                                           - GAGGATAAGC GGGAGATGTG GATGGCTTGT ATTAAGGAGC TGCATGATGT GA - #GCAAGGGC        1200                                                                           - GCCGCTAACA AGTTGGGGGG TGCACTGCAG GCTAAGGCCC GTGCTAAAAA GG - #ATGAACTT        1260                                                                           - AGGAGAAAGA TGATGTATAT GTGCTACAGG AATATAGAGT TCTTTACCAA GA - #ACTCAGCC        1320                                                                           - TTCCCTAAGA CCACCAATGG CTGCAGTCAG GCCATGGCGG CACTGCAGAA CT - #TGCCTCAG        1380                                                                           - TGCTCCCCTG ATGAGATTAT GGCTTATGCC CAGAAAATAT TTAAGATTTT GG - #ATGAGGAG        1440                                                                           - AGAGACAAGG TGCTCACGCA CATTGATCAC ATATTTATGG ATATCCTCAC TA - #CATGTGTG        1500                                                                           - GAAACAATGT GTAATGAGTA CAAGGTCACT AGTGACGCTT GTATGATGAC CA - #TGTACGGG        1560                                                                           - GGCATCTCTC TCTTAAGTGA GTTCTGTCGG GTGCTGTGCT GCTATGTCTT AG - #AGGAGACT        1620                                                                           - AGTGTGATGC TGGCCAAGCG GCCTCTGATA ACCAAGCCTG AGGTTATCAG TG - #TAATGAAG        1680                                                                           - CGCCGCATTG AGGAGATCTG CATGAAGGTC TTTGCCCAGT ACATTCTGGG GG - #CCGATCCT        1740                                                                           - CTGAGAGTCT GCTCTCCTAG TGTGGATGAC CTACGGGCCA TCGCCGAGGA GT - #CAGATGAG        1800                                                                           - GAAGAGGCTA TTGTAGCCTA CACTTTGGCC ACCGCTGGTG TCAGCTCCTC TG - #ATTCTCTG        1860                                                                           - GTGTCACCCC CAGAGTCCCC TGTACCCGCG ACTATCCCTC TGTCCTCAGT AA - #TTGTGGCT        1920                                                                           - GAGAACAGTG ATCAGGAAGA AAGTGAGCAG AGTGATGAGG AAGAGGAGGA GG - #GTGCTCAG        1980                                                                           - GAGGAGCGGG AGGACACTGT GTCTGTCAAG TCTGAGCCAG TGTCTGAGAT AG - #AGGAAGTT        2040                                                                           - GCCCCAGAGG AAGAGGAGGA TGGTGCTGAG GAACCCACCG CCTCTGGAGG TA - #AGAGTACC        2100                                                                           - CACCCTATGG TGACTAGAAG CAAGGCTGAC CAGTAATTTT TATCTCGAGC CC - #GGGAGATC        2160                                                                           - TTAGCTAACT GATTTTTCTG GGAAAAAAAT TATTTAACTT TTCATTAATA GG - #GATTTGAC        2220                                                                           - GTATGTAGCG TACAAAATTA TCGTTCCTGG TATATAGATA AAGAGTCCTA TA - #TATTTGAA        2280                                                                           - AATCGTTACG GCTCGATTAA ACTTTAATGA TTGCATAGTG AATATATCAT TA - #GGATTTAA        2340                                                                           - CTCCTTGACT ATCATGGCGG CGCCAGAAAT TACCATCAAA AGCATTAATA CA - #GTTATGCC        2400                                                                           - GATCGCAGTT AGAACGGTTA TAGCATCCAC CATTTATATC TAAAAATTAG AT - #CAAAGAAT        2460                                                                           - ATGTGACAAA GTCCTAGTTG TATACTGAGA ATTGACGAAA CAATGTTTCT TA - #CATATTTT        2520                                                                           - TTTCTTATTA GTAACTGACT TAATAGTAGG AACTGGAAAG CTAGACTTGA TT - #ATTCTATA        2580                                                                           - AGTATAGATA CCCTTCCAGA TAATGTTCTC TTTGATAAAA GTTCCAGAAA AT - #GTAGAATT        2640                                                                           - TTTTAAAAAG TTATCTTTTG CTATTACCAA GATTGTGTTT AGACGCTTAT TA - #TTAATATG        2700                                                                           #  2739            CCGC CTCTAGATAT GGGGAATTC                                   - (2) INFORMATION FOR SEQ ID NO:60:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 3622 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:60:                                 - GAATTGCGGC CGCTGAATGT TAAATGTTAT ACTTTGGATG AAGCTATAAA TA - #TGCATTGG          60                                                                           - AAAAATAATC CATTTAAAGA AAGGATTCAA ATACTACAAA ACCTAAGCGA TA - #ATATGTTA         120                                                                           - ACTAAGCTTA TTCTTAACGA CGCTTTAAAT ATACACAAAT AAACATAATT TT - #TGTATAAC         180                                                                           - CTAACAAATA ACTAAAACAT AAAAATAATA AAAGGAAATG TAATATCGTA AT - #TATTTTAC         240                                                                           - TCAGGAATGG GGTTAAATAT TTATATCACG TGTATATCTA TACTGTTATC GT - #ATACTCTT         300                                                                           - TACAATTACT ATTACGAATA TGCAAGAGAT AATAAGATTA CGTATTTAAG AG - #AATCTTGT         360                                                                           - CATGATAATT GGGTACGACA TAGTGATAAA TGCTATTTCG CATCGTTACA TA - #AAGTCAGT         420                                                                           - TGGAAAGATG GATTTGACAG ATGTAACTTA ATAGGTGCAA AAATGTTAAA TA - #ACAGCATT         480                                                                           - CTATCGGAAG ATAGGATACC AGTTATATTA TACAAAAATC ACTGGTTGGA TA - #AAACAGAT         540                                                                           - TCTGCAATAT TCGTAAAAGA TGAAGATTAC TGCGAATTTG TAAACTATGA CA - #ATAAAAAG         600                                                                           - CCATTTATCT CAACGACATC GTGTAATTCT TCCATGTTTT ATGTATGTGT TT - #CAGATATT         660                                                                           - ATGAGATTAC TATAAACTTT TTGTATACTT ATATTCCGTA AACTATATTA AT - #CATGAAGA         720                                                                           - AAATGAAAAA GTATAGAAGC TGTTCACGAG CGGTTGTTGA AAACAACAAA AT - #TATACATT         780                                                                           - CAAGATGGCT TACATATACG TCTGTGAGGC TATCATGGAT AATGACAATG CA - #TCTCTAAA         840                                                                           - TAGGTTTTTG GACAATGGAT TCGACCCTAA CACGGAATAT GGTACTCTAC AA - #TCTCCTCT         900                                                                           - TGAAATGGCT GTAATGTTCA AGAATACCGA GGCTATAAAA ATCTTGATGA GG - #TATGGAGC         960                                                                           - TAAACCTGTA GTTACTGAAT GCACAACTTC TTGTCTGCAT GATGCGGTGT TG - #AGAGACGA        1020                                                                           - CTACAAAATA GTGAAAGATC TGTTGAAGAA TAACTATGTA AACAATGTTC TT - #TACAGCGG        1080                                                                           - AGGCTTTACT CCTTTGTGTT TGGCAGCTTA CCTTAACAAA GTTAATTTGG TT - #AAACTTCT        1140                                                                           - ATTGGCTCAT TCGGCGGATG TAGATATTTC AAACACGGAT CGGTTAACTC CT - #CTACATAT        1200                                                                           - AGCCGTATCA AATAAAAATT TAACAATGGT TAAACTTCTA TTGAACAAAG GT - #GCTGATAC        1260                                                                           - TGACTTGCTG GATAACATGG GACGTACTCC TTTAATGATC GCTGTACAAT CT - #GGAAATAT        1320                                                                           - TGAAATATGT AGCACACTAC TTAAAAAAAA TAAAATGTCC AGAACTGGGA AA - #AATTGATC        1380                                                                           - TTGCCAGCTG TAATTCATGG TAGAAAAGAA GTGCTCAGGC TACTTTTCAA CA - #AAGGAGCA        1440                                                                           - GATGTAAACT ACATCTTTGA AAGAAATGGA AAATCATATA CTGTTTTGGA AT - #TGATTAAA        1500                                                                           - GAAAGTTACT CTGAGACACA AAAGAGGTAG CTGAAGTGGT ACTCTCAAAG GT - #ACGTGACT        1560                                                                           - AATTAGCTAT AAAAAGGATC CGGGTTAATT AATTAGTCAT CAGGCAGGGC GA - #GAACGAGA        1620                                                                           - CTATCTGCTC GTTAATTAAT TAGAGCTTCT TTATTCTATA CTTAAAAAGT GA - #AAATAAAT        1680                                                                           - ACAAAGGTTC TTGAGGGTTG TGTTAAATTG AAAGCGAGAA ATAATCATAA AT - #TATTTCAT        1740                                                                           - TATCGCGATA TCCGTTAAGT TTGTATCGTA ATGACGACGT TCCTGCAGAC TA - #TGTTGAGG        1800                                                                           - AAGGAGGTTA ACAGTCAGCT GAGTCTGGGA GACCCGCTGT TTCCAGAGTT GG - #CCGAAGAA        1860                                                                           - TCCCTCAAAA CTTTTGAACA AGTGACCGAG GATTGCAACG AGAACCCCGA GA - #AAGATGTC        1920                                                                           - CTGGCAGAAC TCGTCAAACA GATTAAGGTT CGAGTGGACA TGGTGCGGCA TA - #GAATCAAG        1980                                                                           - GAGCACATGC TGAAAAAATA TACCCAGACG GAAGAGAAAT TCACTGGCGC CT - #TTAATATG        2040                                                                           - ATGGGAGGAT GTTTGCAGAA TGCCTTAGAT ATCTTAGATA AGGTTCATGA GC - #CTTTCGAG        2100                                                                           - GAGATGAAGT GTATTGGGCT AACTATGCAG AGCATGTATG AGAACTACAT TG - #TACCTGAG        2160                                                                           - GATAAGCGGG AGATGTGGAT GGCTTGTATT AAGGAGCTGC ATGATGTGAG CA - #AGGGCGCC        2220                                                                           - GCTAACAAGT TGGGGGGTGC ACTGCAGGCT AAGGCCCGTG CTAAAAAGGA TG - #AACTTAGG        2280                                                                           - AGAAAGATGA TGTATATGTG CTACAGGAAT ATAGAGTTCT TTACCAAGAA CT - #CAGCCTTC        2340                                                                           - CCTAAGACCA CCAATGGCTG CAGTCAGGCC ATGGCGGCAC TGCAGAACTT GC - #CTCAGTGC        2400                                                                           - TCCCCTGATG AGATTATGGC TTATGCCCAG AAAATATTTA AGATTTTGGA TG - #AGGAGAGA        2460                                                                           - GACAAGGTGC TCACGCACAT TGATCACATA TTTATGGATA TCCTCACTAC AT - #GTGTGGAA        2520                                                                           - ACAATGTGTA ATGAGTACAA GGTCACTAGT GACGCTTGTA TGATGACCAT GT - #ACGGGGGC        2580                                                                           - ATCTCTCTCT TAAGTGAGTT CTGTCGGGTG CTGTGCTGCT ATGTCTTAGA GG - #AGACTAGT        2640                                                                           - GTGATGCTGG CCAAGCGGCC TCTGATAACC AAGCCTGAGG TTATCAGTGT AA - #TGAAGCGC        2700                                                                           - CGCATTGAGG AGATCTGCAT GAAGGTCTTT GCCCAGTACA TTCTGGGGGC CG - #ATCCTCTG        2760                                                                           - AGAGTCTGCT CTCCTAGTGT GGATGACCTA CGGGCCATCG CCGAGGAGTC AG - #ATGAGGAA        2820                                                                           - GAGGCTATTG TAGCCTACAC TTTGGCCACC GCTGGTGTCA GCTCCTCTGA TT - #CTCTGGTG        2880                                                                           - TCACCCCCAG AGTCCCCTGT ACCCGCGACT ATCCCTCTGT CCTCAGTAAT TG - #TGGCTGAG        2940                                                                           - AACAGTGATC AGGAAGAAAG TGAGCAGAGT GATGAGGAAG AGGAGGAGGG TG - #CTCAGGAG        3000                                                                           - GAGCGGGAGG ACACTGTGTC TGTCAAGTCT GAGCCAGTGT CTGAGATAGA GG - #AAGTTGCC        3060                                                                           - CCAGAGGAAG AGGAGGATGG TGCTGAGGAA CCCACCGCCT CTGGAGGTAA GA - #GTACCCAC        3120                                                                           - CCTATGGTGA CTAGAAGCAA GGCTGACCAG TAATTTTTAT CTCGAGTCTA GA - #ATCGATCC        3180                                                                           - CGGGTTTTTA TGACTAGTTA ATCACGGCCG CTTATAAAGA TCTAAAATGC AT - #AATTTCTA        3240                                                                           - AATAATGAAA AAAAAGTACA TCATGAGCAA CGCGTTAGTA TATTTTACAA TG - #GAGATTAA        3300                                                                           - CGCTCTATAC CGTTCTATGT TTATTGATTC AGATGATGTT TTAGAAAAGA AA - #GTTATTGA        3360                                                                           - ATATGAAAAC TTTAATGAAG ATGAAGATGA CGACGATGAT TATTGTTGTA AA - #TCTGTTTT        3420                                                                           - AGATGAAGAA GATGACGCGC TAAAGTATAC TATGGTTACA AAGTATAAGT CT - #ATACTACT        3480                                                                           - AATGGCGACT TGTGCAAGAA GGTATAGTAT AGTGAAAATG TTGTTAGATT AT - #GATTATGA        3540                                                                           - AAAACCAAAT AAATCAGATC CATATCTAAA GGTATCTCCT TTGCACATAA TT - #TCATCTAT        3600                                                                           #               3622TGC AG                                                     - (2) INFORMATION FOR SEQ ID NO:61:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 1686 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61:                                 - ATGGAGTCGC GCGGTCGCCG TTGTCCCGAA ATGATATCCG TACTGGGTCC CA - #TTTCGGGG          60                                                                           - CACGTGCTGA AAGCCGTGTT TAGTCGCGGC GACACGCCGG TGCTGCCGCA CG - #AGACGCGA         120                                                                           - CTCCTGCAGA CGGGTATCCA CGTGCGCGTG AGCCAGCCCT CGCTGATCCT GG - #TGTCGCAG         180                                                                           - TACACGCCCG ACTCGACGCC ATGCCACCGC GGCGACAATC AGCTGCAGGT GC - #AGCACACG         240                                                                           - TACTTTACGG GCAGCGAGGT GGAGAACGTG TCGGTCAACG TGCACAACCC CA - #CGGGCCGG         300                                                                           - AGCATCTGCC CCAGCCAAGA GCCCATGTCG ATCTATGTGT ACGCGCTGCC GC - #TCAAGATG         360                                                                           - CTGAACATCC CCAGCATCAA CGTGCACCAC TACCCGTCGG CGGCCGAGCG CA - #AACACCGA         420                                                                           - CACCTGCCCG TAGCTGACGC TGTGATTCAC GCGTCGGGCA AGCAGATGTG GC - #AGGCGCGT         480                                                                           - CTCACGGTCT CGGGACTGGC CTGGACGCGT CAGCAGAACC AGTGGAAAGA GC - #CCGACGTC         540                                                                           - TACTACACGT CAGCGTTCGT GTTTCCCACC AAGGACGTGG CACTGCGGCA CG - #TGGTGTGC         600                                                                           - GCGCACGAGC TGGTTTGCTC CATGGAGAAC ACGCGCGCAA CCAAGATGCA GG - #TGATAGGT         660                                                                           - GACCAGTACG TCAAGGTGTA CCTGGAGTCC TTCTGCGAGG ACGTGCCCTC CG - #GCAAGCTC         720                                                                           - TTTATGCACG TCACGCTGGG CTCTGACGTG GAAGAGGACC TGACGATGAC CC - #GCAACCCG         780                                                                           - CAACCCTTCA TGCGCCCCCA CGAGCGCAAC GGCTTTACGG TGTTGTGTCC CA - #AAAATATG         840                                                                           - ATAATCAAAC CGGGCAAGAT CTCGCACATC ATGCTGGATG TGGCTTTTAC CT - #CACACGAG         900                                                                           - CATTTTGGGC TGCTGTGTCC CAAGAGCATC CCGGGCCTGA GCATCTCAGG TA - #ACCTATTG         960                                                                           - ATGAACGGGC AGCAGATCTT CCTGGAGGTG CAAGCGATAC GCGAGACCGT GG - #AACTGCGT        1020                                                                           - CAGTACGATC CCGTGGCTGC GCTCTTCTTT TTCGATATCG ACTTGCTGCT GC - #AGCGCGGG        1080                                                                           - CCTCAGTACA GCGAACACCC CACCTTCACC AGCCAGTATC GCATCCAGGG CA - #AGCTTGAG        1140                                                                           - TACCGACACA CCTGGGACCG GCACGACGAG GGTGCCGCCC AGGGCGACGA CG - #ACGTCTGG        1200                                                                           - ACCAGCGGAT CGGACTCCGA CGAGGAACTC GTAACCACCG AGCGCAAGAC GC - #CCCGCGTT        1260                                                                           - ACCGGCGGCG GCGCCATGGC GGGCGCCTCC ACTTCCGCGG GCCGCAAACG CA - #AATCAGCA        1320                                                                           - TCCTCGGCGA CGGCGTGCAC GGCGGGCGTT ATGACACGCG GCCGCCTTAA GG - #CCGAGTCC        1380                                                                           - ACCGTCGCGC CCGAAGAGGA CACCGACGAG GATTCCGACA ACGAAATCCA CA - #ATCCGGCC        1440                                                                           - GTGTTCACCT GGCCGCCCTG GCAGGCCGGC ATCCTGGCCC GCAACCTGGT GC - #CCATGGTG        1500                                                                           - GCTACGGTTC AGGGTCAGAA TCTGAAGTAC CAGGAGTTCT TCTGGGACGC CA - #ACGACATC        1560                                                                           - TACCGCATCT TCGCCGAATT GGAAGGCGTA TGGCAGCCCG CTGCGCAACC CA - #AACGTCGC        1620                                                                           - CGCCACCGGC AAGACGCCTT GCCCGGGCCA TGCATCGCCT CGACGCCCAA AA - #AGCACCGA        1680                                                                           #         1686                                                                 - (2) INFORMATION FOR SEQ ID NO:62:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 2745 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62:                                 - GTCGACGATT GTTCATGATG GCAAGATTTA TATATCTGGA GGTTACAACA AT - #AGTAGTGT          60                                                                           - AGTTAATGTA ATATCGAATC TAGTCCTTAG CTATAATCCG ATATATGATG AA - #TGGACCAA         120                                                                           - ATTATCATCA TTAAACATTC CTAGAATTAA TCCCGCTCTA TGGTCAGCGC AT - #AATAAATT         180                                                                           - ATATGTAGGA GGAGGAATAT CTGATGATGT TCGAACTAAT ACATCTGAAA CA - #TACGATAA         240                                                                           - AGAAAAAGAT TGTTGGACAT TGGATAATGG TCACGTGTTA CCACGCAATT AT - #ATAATGTA         300                                                                           - TAAATGCGAA CCGATTAAAC ATAAATATCC ATTGGAAAAA ACACAGTACA CG - #AATGATTT         360                                                                           - TCTAAAGTAT TTGGAAAGTT TTATAGGTAG TTGATAGAAC AAAATACATA AT - #TTTGTAAA         420                                                                           - AATAAATCAC TTTTTATACT AATATTTAAT TAATTAAGCT TGGTACCCTC GA - #AGCTTCTT         480                                                                           - TATTCTATAC TTAAAAAGTG AAAATAAATA CAAAGGTTCT TGAGGGTTGT GT - #TAAATTGA         540                                                                           - AAGCGAGAAA TAATCATAAA TTATTTCATT ATCGCGATAT CCGTTAAGTT TG - #TATCGTAA         600                                                                           - TGGAGTCGCG CGGTCGCCGT TGTCCCGAAA TGATATCCGT ACTGGGTCCC AT - #TTCGGGGC         660                                                                           - ACGTGCTGAA AGCCGTGTTT AGTCGCGGCG ACACGCCGGT GCTGCCGCAC GA - #GACGCGAC         720                                                                           - TCCTGCAGAC GGGTATCCAC GTGCGCGTGA GCCAGCCCTC GCTGATCCTG GT - #GTCGCAGT         780                                                                           - ACACGCCCGA CTCGACGCCA TGCCACCGCG GCGACAATCA GCTGCAGGTG CA - #GCACACGT         840                                                                           - ACTTTACGGG CAGCGAGGTG GAGAACGTGT CGGTCAACGT GCACAACCCC AC - #GGGCCGGA         900                                                                           - GCATCTGCCC CAGCCAAGAG CCCATGTCGA TCTATGTGTA CGCGCTGCCG CT - #CAAGATGC         960                                                                           - TGAACATCCC CAGCATCAAC GTGCACCACT ACCCGTCGGC GGCCGAGCGC AA - #ACACCGAC        1020                                                                           - ACCTGCCCGT AGCTGACGCT GTGATTCACG CGTCGGGCAA GCAGATGTGG CA - #GGCGCGTC        1080                                                                           - TCACGGTCTC GGGACTGGCC TGGACGCGTC AGCAGAACCA GTGGAAAGAG CC - #CGACGTCT        1140                                                                           - ACTACACGTC AGCGTTCGTG TTTCCCACCA AGGACGTGGC ACTGCGGCAC GT - #GGTGTGCG        1200                                                                           - CGCACGAGCT GGTTTGCTCC ATGGAGAACA CGCGCGCAAC CAAGATGCAG GT - #GATAGGTG        1260                                                                           - ACCAGTACGT CAAGGTGTAC CTGGAGTCCT TCTGCGAGGA CGTGCCCTCC GG - #CAAGCTCT        1320                                                                           - TTATGCACGT CACGCTGGGC TCTGACGTGG AAGAGGACCT GACGATGACC CG - #CAACCCGC        1380                                                                           - AACCCTTCAT GCGCCCCCAC GAGCGCAACG GCTTTACGGT GTTGTGTCCC AA - #AAATATGA        1440                                                                           - TAATCAAACC GGGCAAGATC TCGCACATCA TGCTGGATGT GGCTTTTACC TC - #ACACGAGC        1500                                                                           - ATTTTGGGCT GCTGTGTCCC AAGAGCATCC CGGGCCTGAG CATCTCAGGT AA - #CCTATTGA        1560                                                                           - TGAACGGGCA GCAGATCTTC CTGGAGGTGC AAGCGATACG CGAGACCGTG GA - #ACTGCGTC        1620                                                                           - AGTACGATCC CGTGGCTGCG CTCTTCTTTT TCGATATCGA CTTGCTGCTG CA - #GCGCGGGC        1680                                                                           - CTCAGTACAG CGAACACCCC ACCTTCACCA GCCAGTATCG CATCCAGGGC AA - #GCTTGAGT        1740                                                                           - ACCGACACAC CTGGGACCGG CACGACGAGG GTGCCGCCCA GGGCGACGAC GA - #CGTCTGGA        1800                                                                           - CCAGCGGATC GGACTCCGAC GAGGAACTCG TAACCACCGA GCGCAAGACG CC - #CCGCGTTA        1860                                                                           - CCGGCGGCGG CGCCATGGCG GGCGCCTCCA CTTCCGCGGG CCGCAAACGC AA - #ATCAGCAT        1920                                                                           - CCTCGGCGAC GGCGTGCACG GCGGGCGTTA TGACACGCGG CCGCCTTAAG GC - #CGAGTCCA        1980                                                                           - CCGTCGCGCC CGAAGAGGAC ACCGACGAGG ATTCCGACAA CGAAATCCAC AA - #TCCGGCCG        2040                                                                           - TGTTCACCTG GCCGCCCTGG CAGGCCGGCA TCCTGGCCCG CAACCTGGTG CC - #CATGGTGG        2100                                                                           - CTACGGTTCA GGGTCAGAAT CTGAAGTACC AGGAGTTCTT CTGGGACGCC AA - #CGACATCT        2160                                                                           - ACCGCATCTT CGCCGAATTG GAAGGCGTAT GGCAGCCCGC TGCGCAACCC AA - #ACGTCGCC        2220                                                                           - GCCACCGGCA AGACGCCTTG CCCGGGCCAT GCATCGCCTC GACGCCCAAA AA - #GCACCGAG        2280                                                                           - GTTGATTTTT ATGGATCCCC CGGGTAGCTA GCTAATTTTT CTTTTACGTA TT - #ATATATGT        2340                                                                           - AATAAACGTT CACGTAAATA CAAAACAGAG AACAAAGTCT AGATTTTTGA CT - #TACATAAA        2400                                                                           - TGTCTGGGAT AGTAAAATCT ATCATATTGA GCGGACCATC TGGTTCAGGA AA - #GACAGCCA        2460                                                                           - TAGCCAAAAG ACTATGGGAA TATATTTGGA TTTGTGGTGT CCCATACCAC TA - #GATTTCCT        2520                                                                           - CGTCCTATGG AACGAGAAGG TGTCGATTAC CATTACGTTA ACAGAGAGGC CA - #TCTGGAAG        2580                                                                           - GGAATAGCCG CCGGAAACTT TCTAGAACAT ACTGAGTTTT TAGGAAATAT TT - #ACGGAACT        2640                                                                           - TCTAAAACTG CTGTGAATAC AGCGGCTATT AATAATCGTA TTTGTGTGAT GG - #ATTTAAAC        2700                                                                           #                2745TT TAAAAATACT TACCTGCAGA AGCTT                            - (2) INFORMATION FOR SEQ ID NO:63:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 3706 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63:                                 - AAGCTTCTAT CAAAAGTCTT AATGAGTTAG GTGTAGATAG TATAGATATT AC - #TACAAAGG          60                                                                           - TATTCATATT TCCTATCAAT TCTAAAGTAG ATGATATTAA TAACTCAAAG AT - #GATGATAG         120                                                                           - TAGATAATAG ATACGCTCAT ATAATGACTG CAAATTTGGA CGGTTCACAT TT - #TAATCATC         180                                                                           - ACGCGTTCAT AAGTTTCAAC TGCATAGATC AAAATCTCAC TAAAAAGATA GC - #CGATGTAT         240                                                                           - TTGAGAGAGA TTGGACATCT AACTACGCTA AAGAAATTAC AGTTATAAAT AA - #TACATAAT         300                                                                           - GGATTTTGTT ATCATCAGTT ATATTTAACA TAAGTACAAT AAAAAGTATT AA - #ATAAAAAT         360                                                                           - ACTTACTTAC GAAAAAATGT CATTATTACA AAAACTATAT TTTACAGAAC AA - #TCTATAGT         420                                                                           - AGAGTCCTTT AAGAGTTATA ATTTAAAAGA TAACCATAAT GTAATATTTA CC - #ACATCAGA         480                                                                           - TGTTGATACT GTTGTAGTAA TAAATGAAGA TAATGTACTG TTATCTACAA GA - #TTATTATC         540                                                                           - ATTTGATAAA ATTCTGTTTT TTAACTCCTT TAATAACGGT TTATCAAAAT AC - #GAAACTAT         600                                                                           - TAGTGATACA ATATTAGATA TAGATACTCA TAATTATTAT ATACCTAGTT CT - #TCTTCTTT         660                                                                           - GTTAGATATT CTAAAAAAAA GAGCGTGTGA TTTAGAATTA GAAGATCTAA AT - #TATGCGTT         720                                                                           - AATAGGAGAC AATAGTAACT TATATTATAA AGATATGACT TACATGAATA AT - #TGGTTATT         780                                                                           - TACTAAAGGA TTATTAGATT ACAAGTTTGT ATTATTGCGC GATGTAGATA AA - #TGTTACAA         840                                                                           - ACAGTATAAT AAAAAGAATA CTATAATAGA TATAATACAT CGCGATAACA GA - #CAGTATAA         900                                                                           - CATATGGGTT AAAAATGTTA TAGAATACTG TTCTCCTGGC TATATATTAT GG - #TTACATGA         960                                                                           - TCTAAAAGCC GCTGCTGAAG ATGATTGGTT AAGATACGAT AACCGTATAA AC - #GAATTATC        1020                                                                           - TGCGGATAAA TTATACACTT TCGAGTTCAT AGTTATATTA GAAAATAATA TA - #AAACATTT        1080                                                                           - ACGAGTAGGT ACAATAATTG TACATCCAAA CAAGATAATA GCTAATGGTA CA - #TCTAATAA        1140                                                                           - TATACTTACT GATTTTCTAT CTTACGTAGA AGAACTAATA TATCATCATA AT - #TCATCTAT        1200                                                                           - AATATTGGCC GGATATTTTT TAGAATTCTT TGAGACCACT ATTTTATCAG AA - #TTTATTTC        1260                                                                           - TTCATCTTCT GAATGGGTAA TGAATAGTAA CTGTTTAGTA CACCTGAAAA CA - #GGGTATGA        1320                                                                           - AGCTATACTC TTTGATGCTA GTTTATTTTT CCAACTCTCT ACTAAAAGCA AT - #TATGTAAA        1380                                                                           - ATATTGGACA AAGAAAACTT TGCAGTATAA GAACTTTTTT AAAGACGGTA AA - #CAGTTAGC        1440                                                                           - AAAATATATA ATTAAGAAAG ATAGTCAGGT GATAGATAGA GTATGTTATT TA - #CACGCAGC        1500                                                                           - TGTATATAAT CACGTAACTT ACTTAATGGA TACGTTTAAA ATTCCTGGTT TT - #GATTTTAA        1560                                                                           - ATTCTCCGGA ATGATAGATA TACTACTGTT TGGAATATTG CATAAGGATA AT - #GAGAATAT        1620                                                                           - ATTTTATCCG AAACGTGTTT CTGTAACTAA TATAATATCA GAATCTATCT AT - #GCAGATTT        1680                                                                           - TTACTTTATA TCAGATGTTA ATAAATTCAG TAAAAAGATA GAATATAAAA CT - #ATGTTTCC        1740                                                                           - TATACTCGCA GAAAACTACT ATCCAAAAGG AAGGCCCTAT TTTACACATA CA - #TCTAACGA        1800                                                                           - AGATCTTCTG TCTATCTGTT TATGCGAAGT AACAGTTTGT AAAGATATAA AA - #AATCCATT        1860                                                                           - ATTATATTCT AAAAAGGATA TATCAGCAAA ACGATTCATA GGTTTATTTA CA - #TCTGTCGA        1920                                                                           - TATAAATACG GCTGTTGAGT TAAGAGGATA TAAAATAAGA GTAATAGGAT GT - #TTAGAATG        1980                                                                           - GCCTGAAAAG ATAAAAATAT TTAATTCTAA TCCTACATAC ATTAGATTAT TA - #CTAACAGA        2040                                                                           - AAGACGTTTA GATATTCTAC ATTCCTATCT GCTTAAATTT AATATAACAG AG - #GATATAGC        2100                                                                           - TACCAGAGAT GGAGTCAGAA ATAATTTACC TATAATTTCT TTTATCGTCA GT - #TATTGTAG        2160                                                                           - ATCGTATACT TATAAATTAC TAAATTGCCA TATGTACAAT TCGTGTAAGA TA - #ACAAAGTG        2220                                                                           - TAAATATAAT CAGGTAATAT ATAATCCTAT ATAGGAGTAT ATATAATTGA AA - #AAGTAAAA        2280                                                                           - ATAAATCATA TAATAATGAA ACGAAATATC AGTAATAGAC AGGAACTGGC AG - #ATTCTTCT        2340                                                                           - TCTAATGAAG TAAGTACTGC TAAATCTCCA AAATTAGATA AAAATGATAC AG - #CAAATACA        2400                                                                           - GCTTCATTCA ACGAATTACC TTTTAATTTT TTCAGACACA CCTTATTACA AA - #CTAACTAA        2460                                                                           - GTCAGATGAT GAGAAAGTAA ATATAAATTT AACTTATGGG TATAATATAA TA - #AAGATTCA        2520                                                                           - TGATATTAAT AATTTACTTA ACGATGTTAA TAGACTTATT CCATCAACCC CT - #TCAAACCT        2580                                                                           - TTCTGGATAT TATAAAATAC CAGTTAATGA TATTAAAATA GATTGTTTAA GA - #GATGTAAA        2640                                                                           - TAATTATTTG GAGGTAAAGG ATATAAAATT AGTCTATCTT TCACATGGAA AT - #GAATTACC        2700                                                                           - TAATATTAAT AATTATGATA GGAATTTTTT AGGATTTACA GCTGTTATAT GT - #ATCAACAA        2760                                                                           - TACAGGCAGA TCTATGGTTA TGGTAAAACA CTGTAACGGG AAGCAGCATT CT - #ATGGTAAC        2820                                                                           - TGGCCTATGT TTAATAGCCA GATCATTTTA CTCTATAAAC ATTTTACCAC AA - #ATAATAGG        2880                                                                           - ATCCTCTAGA TATTTAATAT TATATCTAAC AACAACAAAA AAATTTAACG AT - #GTATGGCC        2940                                                                           - AGAAGTATTT TCTACTAATA AAGATAAAGA TAGTCTATCT TATCTACAAG AT - #ATGAAAGA        3000                                                                           - AGATAATCAT TTAGTAGTAG CTACTAATAT GGAAAGAAAT GTATACAAAA AC - #GTGGAAGC        3060                                                                           - TTTTATATTA AATAGCATAT TACTAGAAGA TTTAAAATCT AGACTTAGTA TA - #ACAAAACA        3120                                                                           - GTTAAATGCC AATATCGATT CTATATTTCA TCATAACAGT AGTACATTAA TC - #AGTGATAT        3180                                                                           - ACTGAAACGA TCTACAGACT CAACTATGCA AGGAATAAGC AATATGCCAA TT - #ATGTCTAA        3240                                                                           - TATTTTAACT TTAGAACTAA AACGATTCTA CCAATACTAA AAATAGGATA CG - #TGATAGGC        3300                                                                           - TGTTAAAAGC TGCAATAAAT AGTAAGGATG TAGAAGAAAT ACTTTGTTCT AT - #ACCTTCGG        3360                                                                           - AGGAAAGAAC TTTAGAACAA CTTAAGTTTA ATCAAACTTG TATTTATGAA CA - #CTATAAAA        3420                                                                           - AAATTATGGA AGATACAAGT AAAAGAATGG ATGTTGAATG TCGTAGTTTA GA - #ACATAACT        3480                                                                           - ATACGGCTAA CTTATATAAA GTGTACGGAC AAAACGAATA TATGATTACT TA - #TATACTAG        3540                                                                           - CTCTCATAAG TAGGATTAAT AATATTATAG AAACTTTAAA ATATAATCTG GT - #GGGGCTAG        3600                                                                           - ACGAATCTAC AATACGTAAT ATAAATTATA TAATTTCACA AAGAACAAAA AA - #AAATCAGT        3660                                                                           #               3706TAA ACTATATTTT TTACCACTGA CAACAC                           - (2) INFORMATION FOR SEQ ID NO:64:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 3521 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64:                                 - GAGCTCGCGG CCGCCTATCA AAAGTCTTAA TGAGTTAGGT GTAGATAGTA TA - #GATATTAC          60                                                                           - TACAAAGGTA TTCATATTTC CTATCAATTC TAAAGTAGAT GATATTAATA AC - #TCAAAGAT         120                                                                           - GATGATAGTA GATAATAGAT ACGCTCATAT AATGACTGCA AATTTGGACG GT - #TCACATTT         180                                                                           - TAATCATCAC GCGTTCATAA GTTTCAACTG CATAGATCAA AATCTCACTA AA - #AAGATAGC         240                                                                           - CGATGTATTT GAGAGAGATT GGACATCTAA CTACGCTAAA GAAATTACAG TT - #ATAAATAA         300                                                                           - TACATAATGG ATTTTGTTAT CATCAGTTAT ATTTAACATA AGTACAATAA AA - #AGTATTAA         360                                                                           - ATAAAAATAC TTACTTACGA AAAAATGACT AATTAGCTAT AAAAACCCAA CA - #AAAACTAA         420                                                                           - TCAGCTATCG GGGTTAATTA ATTAGTTATT AGACAAGGTG AAAACGAAAC TA - #TTTGTAGC         480                                                                           - TTAATTAATT AGAGCTTCTT TATTCTATAC TTAAAAAGTG AAAATAAATA CA - #AAGGTTCT         540                                                                           - TGAGGGTTGT GTTAAATTGA AAGCGAGAAA TAATCATAAA TTATTTCATT AT - #CGCGATAT         600                                                                           - CCGTTAAGTT TGTATCGTAA TGGAGTCGCG CGGTCGCCGT TGTCCCGAAA TG - #ATATCCGT         660                                                                           - ACTGGGTCCC ATTTCGGGGC ACGTGCTGAA AGCCGTGTTT AGTCGCGGCG AC - #ACGCCGGT         720                                                                           - GCTGCCGCAC GAGACGCGAC TCCTGCAGAC GGGTATCCAC GTGCGCGTGA GC - #CAGCCCTC         780                                                                           - GCTGATCCTG GTGTCGCAGT ACACGCCCGA CTCGACGCCA TGCCACCGCG GC - #GACAATCA         840                                                                           - GCTGCAGGTG CAGCACACGT ACTTTACGGG CAGCGAGGTG GAGAACGTGT CG - #GTCAACGT         900                                                                           - GCACAACCCC ACGGGCCGGA GCATCTGCCC CAGCCAAGAG CCCATGTCGA TC - #TATGTGTA         960                                                                           - CGCGCTGCCG CTCAAGATGC TGAACATCCC CAGCATCAAC GTGCACCACT AC - #CCGTCGGC        1020                                                                           - GGCCGAGCGC AAACACCGAC ACCTGCCCGT AGCTGACGCT GTGATTCACG CG - #TCGGGCAA        1080                                                                           - GCAGATGTGG CAGGCGCGTC TCACGGTCTC GGGACTGGCC TGGACGCGTC AG - #CAGAACCA        1140                                                                           - GTGGAAAGAG CCCGACGTCT ACTACACGTC AGCGTTCGTG TTTCCCACCA AG - #GACGTGGC        1200                                                                           - ACTGCGGCAC GTGGTGTGCG CGCACGAGCT GGTTTGCTCC ATGGAGAACA CG - #CGCGCAAC        1260                                                                           - CAAGATGCAG GTGATAGGTG ACCAGTACGT CAAGGTGTAC CTGGAGTCCT TC - #TGCGAGGA        1320                                                                           - CGTGCCCTCC GGCAAGCTCT TTATGCACGT CACGCTGGGC TCTGACGTGG AA - #GAGGACCT        1380                                                                           - GACGATGACC CGCAACCCGC AACCCTTCAT GCGCCCCCAC GAGCGCAACG GC - #TTTACGGT        1440                                                                           - GTTGTGTCCC AAAAATATGA TAATCAAACC GGGCAAGATC TCGCACATCA TG - #CTGGATGT        1500                                                                           - GGCTTTTACC TCACACGAGC ATTTTGGGCT GCTGTGTCCC AAGAGCATCC CG - #GGCCTGAG        1560                                                                           - CATCTCAGGT AACCTATTGA TGAACGGGCA GCAGATCTTC CTGGAGGTGC AA - #GCGATACG        1620                                                                           - CGAGACCGTG GAACTGCGTC AGTACGATCC CGTGGCTGCG CTCTTCTTTT TC - #GATATCGA        1680                                                                           - CTTGCTGCTG CAGCGCGGGC CTCAGTACAG CGAACACCCC ACCTTCACCA GC - #CAGTATCG        1740                                                                           - CATCCAGGGC AAGCTTGAGT ACCGACACAC CTGGGACCGG CACGACGAGG GT - #GCCGCCCA        1800                                                                           - GGGCGACGAC GACGTCTGGA CCAGCGGATC GGACTCCGAC GAGGAACTCG TA - #ACCACCGA        1860                                                                           - GCGCAAGACG CCCCGCGTTA CCGGCGGCGG CGCCATGGCG GGCGCCTCCA CT - #TCCGCGGG        1920                                                                           - CCGCAAACGC AAATCAGCAT CCTCGGCGAC GGCGTGCACG GCGGGCGTTA TG - #ACACGCGG        1980                                                                           - CCGCCTTAAG GCCGAGTCCA CCGTCGCGCC CGAAGAGGAC ACCGACGAGG AT - #TCCGACAA        2040                                                                           - CGAAATCCAC AATCCGGCCG TGTTCACCTG GCCGCCCTGG CAGGCCGGCA TC - #CTGGCCCG        2100                                                                           - CAACCTGGTG CCCATGGTGG CTACGGTTCA GGGTCAGAAT CTGAAGTACC AG - #GAGTTCTT        2160                                                                           - CTGGGACGCC AACGACATCT ACCGCATCTT CGCCGAATTG GAAGGCGTAT GG - #CAGCCCGC        2220                                                                           - TGCGCAACCC AAACGTCGCC GCCACCGGCA AGACGCCTTG CCCGGGCCAT GC - #ATCGCCTC        2280                                                                           - GACGCCCAAA AAGCACCGAG GTTGATTTTT ATGGATCCGG TACCCTCGAG GA - #ATTCTTTT        2340                                                                           - TATTGATTAA CTAGTCAAAT GAGTATATAT AATTGAAAAA GTAAAATATA AA - #TCATATAA        2400                                                                           - TAATGAAACG AAATATCAGT AATAGACAGG AACTGGCAGA TTCTTCTTCT AA - #TGAAGTAA        2460                                                                           - GTACTGCTAA ATCTCCAAAA TTAGATAAAA ATGATACAGC AAATACAGCT TC - #ATTCAACG        2520                                                                           - AATTACCTTT TAATTTTTTC AGACACACCT TATTACAAAC TAACTAAGTC AG - #ATGATGAG        2580                                                                           - AAAGTAAATA TAAATTTAAC TTATGGGTAT AATATAATAA AGATTCATGA TA - #TTAATAAT        2640                                                                           - TTACTTAACG ATGTTAATAG ACTTATTCCA TCAACCCCTT CAAACCTTTC TG - #GATATTAT        2700                                                                           - AAAATACCAG TTAATGATAT TAAAATAGAT TGTTTAAGAG ATGTAAATAA TT - #ATTTGGAG        2760                                                                           - GTAAAGGATA TAAAATTAGT CTATCTTTCA CATGGAAATG AATTACCTAA TA - #TTAATAAT        2820                                                                           - TATGATAGGA ATTTTTTAGG ATTTACAGCT GTTATATGTA TCAACAATAC AG - #GCAGATCT        2880                                                                           - ATGGTTATGG TAAAACACTG TAACGGGAAG CAGCATTCTA TGGTAACTGG CC - #TATGTTTA        2940                                                                           - ATAGCCAGAT CATTTTACTC TATAAACATT TTACCACAAA TAATAGGATC CT - #CTAGATAT        3000                                                                           - TTAATATTAT ATCTAACAAC AACAAAAAAA TTTAACGATG TATGGCCAGA AG - #TATTTTCT        3060                                                                           - ACTAATAAAG ATAAAGATAG TCTATCTTAT CTACAAGATA TGAAAGAAGA TA - #ATCATTTA        3120                                                                           - GTAGTAGCTA CTAATATGGA AAGAAATGTA TACAAAAACG TGGAAGCTTT TA - #TATTAAAT        3180                                                                           - AGCATATTAC TAGAAGATTT AAAATCTAGA CTTAGTATAA CAAAACAGTT AA - #ATGCCAAT        3240                                                                           - ATCGATTCTA TATTTCATCA TAACAGTAGT ACATTAATCA GTGATATACT GA - #AACGATCT        3300                                                                           - ACAGACTCAA CTATGCAAGG AATAAGCAAT ATGCCAATTA TGTCTAATAT TT - #TAACTTTA        3360                                                                           - GAACTAAAAC GTTCTACCAA TACTAAAAAT AGGATACGTG ATAGGCTGTT AA - #AAGCTGCA        3420                                                                           - ATAAATAGTA AGGATGTAGA AGAAATACTT TGTTCTATAC CTTCGGAGGA AA - #GAACTTTA        3480                                                                           # 3521             ATCA AACTTGTATT TATGAAGGTA C                                - (2) INFORMATION FOR SEQ ID NO:65:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 2160 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65:                                 - AAGACTAATT TGTAAACCAT CTTACTCAAA ATATGTAACA ATAGTACGAT GC - #AATGAGTA          60                                                                           - AGACAATAGG AAATCTATCT TATATACACA TAATTATTCT ATCAATTTTA CC - #AATTAGTT         120                                                                           - AGTGTAATGT TATAAAAACT AATTAATCAC TCGAGCCCCC TCGAAGCTTC TT - #TATTCTAT         180                                                                           - ACTTAAAAAG TGAAAATAAA TACAAAGGTT CTTGAGGGTT GTGTTAAATT GA - #AAGCGAGA         240                                                                           - AATAATCATA AATTATTTCA TTATCGCGAT ATCCGTTAAG TTTGTATCGT AA - #TGGAGTCG         300                                                                           - CGCGGTCGCC GTTGTCCCGA AATGATATCC GTACTGGGTC CCATTTCGGG GC - #ACGTGCTG         360                                                                           - AAAGCCGTGT TTAGTCGCGG CGACACGCCG GTGCTGCCGC ACGAGACGCG AC - #TCCTGCAG         420                                                                           - ACGGGTATCC ACGTGCGCGT GAGCCAGCCC TCGCTGATCC TGGTGTCGCA GT - #ACACGCCC         480                                                                           - GACTCGACGC CATGCCACCG CGGCGACAAT CAGCTGCAGG TGCAGCACAC GT - #ACTTTACG         540                                                                           - GGCAGCGAGG TGGAGAACGT GTCGGTCAAC GTGCACAACC CCACGGGCCG GA - #GCATCTGC         600                                                                           - CCCAGCCAAG AGCCCATGTC GATCTATGTG TACGCGCTGC CGCTCAAGAT GC - #TGAACATC         660                                                                           - CCCAGCATCA ACGTGCACCA CTACCCGTCG GCGGCCGAGC GCAAACACCG AC - #ACCTGCCC         720                                                                           - GTAGCTGACG CTGTGATTCA CGCGTCGGGC AAGCAGATGT GGCAGGCGCG TC - #TCACGGTC         780                                                                           - TCGGGACTGG CCTGGACGCG TCAGCAGAAC CAGTGGAAAG AGCCCGACGT CT - #ACTACACG         840                                                                           - TCAGCGTTCG TGTTTCCCAC CAAGGACGTG GCACTGCGGC ACGTGGTGTG CG - #CGCACGAG         900                                                                           - CTGGTTTGCT CCATGGAGAA CACGCGCGCA ACCAAGATGC AGGTGATAGG TG - #ACCAGTAC         960                                                                           - GTCAAGGTGT ACCTGGAGTC CTTCTGCGAG GACGTGCCCT CCGGCAAGCT CT - #TTATGCAC        1020                                                                           - GTCACGCTGG GCTCTGACGT GGAAGAGGAC CTGACGATGA CCCGCAACCC GC - #AACCCTTC        1080                                                                           - ATGCGCCCCC ACGAGCGCAA CGGCTTTACG GTGTTGTGTC CCAAAAATAT GA - #TAATCAAA        1140                                                                           - CCGGGCAAGA TCTCGCACAT CATGCTGGAT GTGGCTTTTA CCTCACACGA GC - #ATTTTGGG        1200                                                                           - CTGCTGTGTC CCAAGAGCAT CCCGGGCCTG AGCATCTCAG GTAACCTATT GA - #TGAACGGG        1260                                                                           - CAGCAGATCT TCCTGGAGGT GCAAGCGATA CGCGAGACCG TGGAACTGCG TC - #AGTACGAT        1320                                                                           - CCCGTGGCTG CGCTCTTCTT TTTCGATATC GACTTGCTGC TGCAGCGCGG GC - #CTCAGTAC        1380                                                                           - AGCGAACACC CCACCTTCAC CAGCCAGTAT CGCATCCAGG GCAAGCTTGA GT - #ACCGACAC        1440                                                                           - ACCTGGGACC GGCACGACGA GGGTGCCGCC CAGGGCGACG ACGACGTCTG GA - #CCAGCGGA        1500                                                                           - TCGGACTCCG ACGAGGAACT CGTAACCACC GAGCGCAAGA CGCCCCGCGT TA - #CCGGCGGC        1560                                                                           - GGCGCCATGG CGGGCGCCTC CACTTCCGCG GGCCGCAAAC GCAAATCAGC AT - #CCTCGGCG        1620                                                                           - ACGGCGTGCA CGGCGGGCGT TATGACACGC GGCCGCCTTA AGGCCGAGTC CA - #CCGTCGCG        1680                                                                           - CCCGAAGAGG ACACCGACGA GGATTCCGAC AACGAAATCC ACAATCCGGC CG - #TGTTCACC        1740                                                                           - TGGCCGCCCT GGCAGGCCGG CATCCTGGCC CGCAACCTGG TGCCCATGGT GG - #CTACGGTT        1800                                                                           - CAGGGTCAGA ATCTGAAGTA CCAGGAGTTC TTCTGGGACG CCAACGACAT CT - #ACCGCATC        1860                                                                           - TTCGCCGAAT TGGAAGGCGT ATGGCAGCCC GCTGCGCAAC CCAAACGTCG CC - #GCCACCGG        1920                                                                           - CAAGACGCCT TGCCCGGGCC ATGCATCGCC TCGACGCCCA AAAAGCACCG AG - #GTTGATTT        1980                                                                           - TTATGGATCC TCGCGACTGC AGGGTACCTG AGTAGCTAAT TTTTAAACAA AA - #ATGTGGGA        2040                                                                           - GAATCTAATT AGTTTTTCTT TACACAATTG ACGTACATGA GTCTGAGTTC CT - #TGTTTTTG        2100                                                                           - CTAATTATTT CATCCAATTT ATTATTCTTG ACGATATCGA GATCTTTTGT AT - #AGGAGTCA        2160                                                                           - (2) INFORMATION FOR SEQ ID NO:66:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 3141 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66:                                 - ATGAGTTTGC AGTTTATCGG TCTACAGCGG CGCGATGTGG TGGCCCTGGT CA - #ACTTTCTG          60                                                                           - CGCCATCTCA CGCAAAAGCC CGACGTGGAT CTCGAGGCAC ACCCCAAGAT CC - #TGAAAAAA         120                                                                           - TGTGGCGAAA AACGCCTGCA CCGGCGTACG GTGCTGTTCA ACGAGCTCAT GC - #TTTGGTTG         180                                                                           - GGATACTACC GCGAGCTGCG TTTCCACAAC CCCGACCTCT CCTCGGTTCT CG - #AGGAGTTC         240                                                                           - GAGGTGCGTT GCGCGGCCGT GGCGCGTCGC GGCTACACTT ACCCGTTCGG TG - #ATCGTGGT         300                                                                           - AAGGCGCGTG ACCACCTGGC TGTGCTAGAC CGTACCGAAT TCGATACGGA CG - #TACGCCAC         360                                                                           - GATGCTGAGA TTGTGGAGCG CGCGCTCGTA AGCGCGGTCA TTCTGGCCAA GA - #TGTCGGTG         420                                                                           - CGCGAGACGC TGGTCACAGC CATCGGCCAG ACGGAACCCA TCGCTTTTGT GC - #ACCTCAAG         480                                                                           - GATACGGAGG TGCAGCGCAT TGAAGAAAAC CTGGAGGGTG TGCGCCGTAA CA - #TGTTCTGC         540                                                                           - GTGAAACCGC TCGACCTTAA CCTGGACCGG CACGCCAACA CGGCGCTGGT CA - #ACGCCGTC         600                                                                           - AACAAGCTCG TGTACACGGG CCGTCTCATC ATGAACGTGC GCAGGTCTTG GG - #AGGAGCTG         660                                                                           - GAGCGCAAAT GTCTGGCGCG CATTCAGGAG CGCTGCAAGC TGCTGGTCAA GG - #AGCTGCGC         720                                                                           - ATGTGCCTTT CCTTTGATTC CAACTACTGT CGCAATATCC TCAAACACGC CG - #TGGAAAAC         780                                                                           - GGTGACTCGG CCGACACGCT GCTGGAGCTG CTCATCGAGG ACTTTGACAT CT - #ACGTGGAC         840                                                                           - AGCTTCCCGC AGTCGGCGCA CACCTTTTTG GGCGCGCGCC CGCCGTCGTT GG - #AGTTTGAC         900                                                                           - GATGACGCCA ATCTCCTCTC GCTCGGCGGC GGTTCAGCCT TCTCGTCGGT AC - #CCAAGAAA         960                                                                           - CATGTCCCCA CGCAGCCGCT GGACGGCTGG AGCTGGATCG CCAGTCCCTG GA - #AGGGACAC        1020                                                                           - AAACCGTTCC GCTTCGAGGC CCATGGTTCT CTGGCACCGG CCGCCGACGC CC - #ACGCCGCC        1080                                                                           - CGTTCGGCGC GCGTCGGCTA TTACGACGAA GAGGAAAAGC GTCGCGAGCG GC - #AGAAACGG        1140                                                                           - GTGGACGACG AGGTGGTGCA GCGTGAGAAA CAGCAGCTGA AGGCTTGGGA GG - #AGAGGCAG        1200                                                                           - CAGAACCTGC AGCAACGTCA GCAGCAACCG CCGCCCCCGA CACGTAAACC GG - #GCGCCTCC        1260                                                                           - CGGAGGCTCT TTGGCTCCAG TGCCGATGAG GACGACGACG ATGATGATGA CG - #AGAAAAAC        1320                                                                           - ATCTTTACGC CCATCAAGAA ACCGGGAACT AGCGGCAAGG GCGCCGCTAG TG - #GCAACGGT        1380                                                                           - GTTTCCAGCA TTTTCAGCGG CATGTTATCC TCGGGCAGTC AGAAACCGAC CA - #GCGGTCCC        1440                                                                           - TTGAACATCC CGCAGCAACA ACAGCGTCAC GCGGCTTTCA GTCTCGTCTC CC - #CGCAGGTA        1500                                                                           - ACCAAGGCCA GCCCGGGAAG GGTCCGTCGG GACAGCGCGT GGGACGTGAG GC - #CGCTCACG        1560                                                                           - GAGACAAGAG GGGATCTTTT CTCGGGCGAC GAGGATTCCG ACAGCTCGGA TG - #GCTATCCC        1620                                                                           - CCCAACCGTC AAGATCCGCG TTTCACCGAC ACGCTGGTGG ACATCACGGA TA - #CCGAGACG        1680                                                                           - AGCGCCAAAC CGCCCGTCAC CACCGCGTAC AAGTTCGAGC AACCGACGTT GA - #CGTTCGGC        1740                                                                           - GCCGGAGTTA ACGTCCCTGC TGGCGCCGGC GCTGCCATCC TCACGCCGAC GC - #CTGTCAAT        1800                                                                           - CCTTCCACGG CCCCCGCTCC GGCCCCGACA CCTACCTTCG CGGGTACCCA AA - #CCCCGGTC        1860                                                                           - AACGGTAACT CGCCCTGGGC TCCGACGGCG CCGTTGCCCG GGGATATGAA CC - #CCGCCAAC        1920                                                                           - TGGCCGCGCG AACGCGCGTG GGCCCTCAAG AATCCTCACC TGGCTTACAA TC - #CCTTCAGG        1980                                                                           - ATGCCTACGA CTTCCACGAC TTCTCAAAAC AACGTGTCCA CCACCCCTCG GA - #GGCCGTCG        2040                                                                           - ACTCCACGCG CCGCGGTGAC ACAAACAGCG TCTCAGAACG CCGCTGATGA GG - #TTTGGGCT        2100                                                                           - TTAAGGGACC AAACTGCAGA GTCACCGGTC GAAGACAGCG AGGAGGAAGA CG - #ACGACTCC        2160                                                                           - TCGGACACCG GCTCCGTCGT CAGCCTGGGA CACACAACAC CGTCGTCCGA TT - #ACAACGAC        2220                                                                           - GTCATTTCGC CTCCCAGTCA GACGCCCGAG CAGTCGACGC CGTCCAGAAT AC - #GTAAAGCT        2280                                                                           - AAGTTATCGT CTCCAATGAC GACGACATCC ACGAGCCAGA AACCGGTGCT GG - #GCAAGCGA        2340                                                                           - GTCGCGACGC CGCACGCGTC CGCCCGAGCG CAGACGGTGA CGTCGACACC GG - #TTCAGGGA        2400                                                                           - AGGGTAGAGA AACAGGTATC GGGCACGCCG TCGACGGTAC CCGCCACGCT GT - #TGCAACCT        2460                                                                           - CAACCGGCTT CGTCTAAAAC AACGTCATCA AGGAACGTGA CTTCTGGCGC GA - #GAACCTCT        2520                                                                           - TCCGCTTCGG CTCGACAGCC GTCAGCCTCG GCGTCCGTTT TGTCGCCCAC GG - #AGGATGAT        2580                                                                           - GTCGTGTCCC CCGTCACGTC GCCGCTGTCC ATGCTTTCGT CAGCCTCTCC GT - #CCCCGGCC        2640                                                                           - AAGAGTGCCC CTCCGTCTCC GGTGAAAGGT CGGGGCAGCC GCGTCGGTGT TC - #CTTCTTTG        2700                                                                           - AAACCTACTT TGGGCGGCAA GGCGGTGGTA GGTCGACCGC CCTCGGTCCC CG - #TGAGCGGT        2760                                                                           - AGCGCGCCGG GTCGCCTGTC CGGCACCAGC CGGGCCGCCT CGACCACGCC GA - #CGTATCCC        2820                                                                           - GCGGTAACCA CCGTTTACCC ACCGTCGTCT ACGGCCAAAA GCAGCGTATC GA - #ATGCGCCG        2880                                                                           - CCTGTGGCCT CCCCCTCCAT CCTGAAACCG GGGGCGAGCG CGGCTTTGCA AT - #CACGCCGC        2940                                                                           - TCGACGGGGA CCGCCGCCGT AGGTTCCCCC GTCAAGAGCA CGACGGGCAT GA - #AAACGGTG        3000                                                                           - GCTTTCGACC TATCGTCGCC CCAGAAGAGC GGTACGGGGC CGCAACCGGG TT - #CTGCCGGC        3060                                                                           - ATGGGGGGCG CCAAAACGCC GTCGGACGCC GTGCAGAACA TCCTCCAAAA GA - #TCGAGAAG        3120                                                                           #                3141TA G                                                      - (2) INFORMATION FOR SEQ ID NO:67:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 4075 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67:                                 - AAGCTTGCGG CCGCTCATTA GACAAGCGAA TGAGGGACGA AAACGTGGAG GA - #GGTATTAA          60                                                                           - GTTTGGAGAA ATGGAGAGAG ACTGTTTAAT AGCGCATGGC GCAGCCAATA CT - #ATTACAGA         120                                                                           - AGTTTTGAAA GATTCGGAAG AAGATTATCA AGATGTGTAT GTTTGTGAAA AT - #TGTGGAGA         180                                                                           - CATAGCAGCA CAAATCAAGG GTATTAATAC ATGTCTTAGA TGTTCAAAAC TT - #AATCTCTC         240                                                                           - TCCTCTCTTA ACAAAAATTG ATACCACGCA CGTATCTAAA GTATTTCTTA CT - #CAAATGAA         300                                                                           - CGCCAGAGGC GTAAAAGTCA AATTAGATTT CGAACGAAGG CCTCCTTCGT TT - #TATAAACC         360                                                                           - ATTAGATAAA GTTGATCTCA AGCCGTCTTT TCTGGTGTAA TAAAAATTAA TT - #AATTACTC         420                                                                           - GAGCCCCTAG CAATAAAAAC TATTCCTCCG TGTTCTTAAT CTTCTCGATC TT - #TTGGAGGA         480                                                                           - TGTTCTGCAC GGCGTCCGAC GGCGTTTTGG CGCCCCCCAT GCCGGCAGAA CC - #CGGTTGCG         540                                                                           - GCCCCGTACC GCTCTTCTGG GGCGACGATA GGTCGAAAGC CACCGTTTTC AT - #GCCCGTCG         600                                                                           - TGCTCTTGAC GGGGGAACCT ACGGCGGCGG TCCCCGTCGA GCGGCGTGAT TG - #CAAAGCCG         660                                                                           - CGCTCGCCCC CGGTTTCAGG ATGGAGGGGG AGGCCACAGG CGGCGCATTC GA - #TACGCTGC         720                                                                           - TTTTGGCCGT AGACGACGGT GGGTAAACGG TGGTTACCGC GGGATACGTC GG - #CGTGGTCG         780                                                                           - AGGCGGCCCG GCTGGTGCCG GACAGGCGAC CCGGCGCGCT ACCGCTCACG GG - #TACCGAGG         840                                                                           - GCGGTCGACC TACCACCGCC TTGCCGCCCA AAGTAGGTTT CAAAGAAGGA AC - #ACCGACGC         900                                                                           - GGCTGCCCCG ACCTTTCACC GGAGACGGAG GGGCACTCTT GGCCGGGGAC GG - #AGAGGCTG         960                                                                           - ACGAAAGCAT GGACAGCGGC GACGTGACGG GGGACACGAC ATCATCCTCC GT - #GGGCGACA        1020                                                                           - AAACGGACGC CGAGGCTGAC GGCTGTCGAG CCGAAGCGGA AGAGGTTCTC GC - #GCCAGAAG        1080                                                                           - TCACGTTCCT TGATGACGTT GTTTTAGACG AAGCCGGTTG AGGTTGCAAC AG - #CGTGGCGG        1140                                                                           - GTACCGTCGA CGGCGTGCCC GATACCTGTT TCTCTACCCT TCCCTGAACC GG - #TGTCGACG        1200                                                                           - TCACCGTCTG CGCTCGGGCG GACGCGTGCG GCGTCGCGAC TCGCTTGCCC AG - #CACCGGTT        1260                                                                           - TCTGGCTCGT GGATGTCGTC GTCATTGGAG ACGATAACTT AGCTTTACGT AT - #TCTGGACG        1320                                                                           - GCGTCGACTG CTCGGGCGTC TGACTGGGAG GCGAAATGAC GTCGTTGTAA TC - #GGACGACG        1380                                                                           - GTGTTGTGTG TCCCAGGCTG ACGACGGAGC CGGTGTCCGA GGAGTCGTCG TC - #TTCCTCCT        1440                                                                           - CGCTGTCTTC GACCGGTGAC TCTGCAGTTT GGTCCCTTAA AGCCCAAACC TC - #ATCAGCGG        1500                                                                           - CGTTCTGAGA CGCTGTTTGT GTCACCGCGG CGCGTGGAGT CGACGGCCTC CG - #AGGGGTGG        1560                                                                           - TGGACACGTT GTTTTGAGAA GTCGTGGAAG TCGTAGGCAT CCTGAAGGGA TT - #GTAAGCCA        1620                                                                           - GGTGAGGATT CTTGAGGGCC CACGCGCGTT CGCGCGGCCA GTTGGCGGGG TT - #CATATCCC        1680                                                                           - CGGGCAACGG CGCCGTCGGA GCCCAGGGCG AGTTACCGTT GACCGGGGTT TG - #GGTACCCG        1740                                                                           - CGAAGGTAGG TGTCGGGGCC GGAGCGGGGG CCGTGGAAGG ATTGACAGGC GT - #CGGCGTGA        1800                                                                           - GGATGGCAGC GCCGGCGCCA GCAGGGACGT TAACTCCGGC GCCGAACGTC AA - #CGTCGGTT        1860                                                                           - GCTCGAACTT GTACGCGGTG GTGACGGGCG GTTTGGCGCT CGTCTCGGTA TC - #CGTGATGT        1920                                                                           - CCACCAGCGT GTCGGTGAAA CGCGGATCTT GACGGTTGGG GGGATAGCCA TC - #CGAGCTGT        1980                                                                           - CGGAATCCTC GTCGCCCGAG AAAAGATCCC CTCTTGTCTC CGTGAGCGGC CT - #CACGTCCC        2040                                                                           - ACGCGCTGTC CCGACGGACC CTTCCCGGGC TGGCCTTGGT TACCTGCGGG GA - #GACGAGAC        2100                                                                           - TGAAAGCCGC GTGACGCTGT TGTTGCTGCG GGATGTTCAA GGGACCGCTG GT - #CGGTTTCT        2160                                                                           - GACTGCCCGA GGATAACATG CCGCTGAAAA TGCTGGAAAC ACCGTTGCCA CT - #AGCGGCGC        2220                                                                           - CCTTGCCGCT AGTTCCCGGT TTCTTGATGG GCGTAAAGAT GTTTTTCTCG TC - #ATCATCAT        2280                                                                           - CGTCGTCGTC CTCATCGGCA CTGGAGCCAA AGAGCCTCCG GGAGGCGCCC GG - #TTTACGTG        2340                                                                           - TCGGGGGCGG CGGTTGCTGC TGACGTTGCT GCAGGTTCTG CTGCCTCTCC TC - #CCAAGCCT        2400                                                                           - TCAGCTGCTG TTTCTCACGC TGCACCACCT CGTCGTCCAC CCGTTTCTGC CG - #CTCGCGAC        2460                                                                           - GCTTTTCCTC TTCGTCGTAA TAGCCGACGC GCGCCGAACG GGCGGCGTGG GC - #GTCGGCGG        2520                                                                           - CCGGTGCCAG AGAACCATGG GCCTCGAAGC GGAACGGTTT GTGTCCCTTC CA - #GGGACTGG        2580                                                                           - CGATCCAGCT CCAGCCGTCC AGCGGCTGCG TGGGGACATG TTTCTTGGGT AC - #CGACGAGA        2640                                                                           - AGGCTGAACC GCCGCCGAGC GAGAGGAGAT TGGCGTCATC GTCAAACTCC AA - #CGACGGCG        2700                                                                           - GGCGCGCGCC CAAAAAGGTG TGCGCCGACT GCGGGAAGCT GTCCACGTAG AT - #GTCAAAGT        2760                                                                           - CCTCGATGAG CAGCTCCAGC AGCGTGTCGG CCGAGTCACC GTTTTCCACG GC - #GTGTTTGA        2820                                                                           - GGATATTGCG ACAGTAGTTG GAATCAAAGG AAAGGCACAT GCGCAGCTCC TT - #GACCAGCA        2880                                                                           - GCTTGCAGCG CTCCTGAATG CGCGCCAGAC ATTTGCGCTC CAGCTCCTCC CA - #AGACCTGC        2940                                                                           - GCACGTTCAT GATGAGACGG CCCGTGTACA CGAGCTTGTT GACGGCGTTG AC - #CAGCGCCG        3000                                                                           - TGTTGGCGTG CCGGTCCAGG TTAAGGTCGA GCGGTTTCAC GCAGAACATG TT - #ACGGCGCA        3060                                                                           - CACCCTCCAG GTTTTCTTCA ATGCGCTGCA CCTCCGTATC CTTGAGGTGC AC - #AAAAGCGA        3120                                                                           - GTGGGTTCCG TCTGGCCGAT GGCTGTGACC AGCGTCTCGC GCACCGACAT CT - #TGGCCAGA        3180                                                                           - ATGACCGCGC TTACGAGCGC GCGCTCCACA ATCTCAGCAT CGTGGCGTAC GT - #CCGTATCG        3240                                                                           - AATTCGGTAC GGTCTAGCAC AGCCAGGTGG TCACGCGCCT TACCACGATC AC - #CGAACGGG        3300                                                                           - TAAGTGTAGC CGCGACGCGC CACGGCCGCG CAACGCACCT CGAACTCCTC GA - #GAACCGAG        3360                                                                           - GAGAGGTCGG GGTTGTGGAA ACGCAGCTCG CGGTAGTATC CCAACCAAAG CA - #TGAGCTCG        3420                                                                           - TTGAACAGCA CCGTAGCCGG TGCAGGCGTT TTTCGCCACA TTTTTTCAGG AT - #CTTGGGGT        3480                                                                           - GTGCCTCGAG ATCCACGTCG GGCTTTTGCG TGAGATGGCG CAGAAAGTTG AC - #CAGGGCCA        3540                                                                           - CCACATCGCG CCGCTGTAGA CCGATAAACT GCAAACTCAT TTTATATTGT AA - #TTATATAT        3600                                                                           - TTTCAATTTT GAAATCCCAA AATATTATCA TATCTTCCCA ATAAAGCTAG GG - #GAGATCTA        3660                                                                           - ATTTAATTTA ATTTATATAA CTTATTTTTT GAATATACTT TTAATTAACA AA - #AGAGTTAA        3720                                                                           - GTTACTCATA TGGACGCCGT CCAGTCTGAA CATCAATCTT TTTAGCCAGA GA - #TATCATAG        3780                                                                           - CCGCTCTTAG AGTTTCAGCG TGATTTTCCA ACCTAAATAG AACTTCATCG TT - #GCGTTTAC        3840                                                                           - AACACTTTTC TATTTGTTCA AACTTTGTTG TTACATTAGT AATCTTTTTT TC - #CAAATTAG        3900                                                                           - TTAGCCGTTG TTTGAGAGTT TCCTCATTGT CGTCTTCATC GGCTTTAACA AT - #TGCTTCGC        3960                                                                           - GTTTAGCCTC CTGGCTGTTC TTATCAGCCT TTGTAGAAAA AAATTCAGTT GC - #TGGAATTG        4020                                                                           - CAAGATCGTC ATCTCCGGGG AAAAGAGTTC CGTCCATTTA AAGCCGCGGG AA - #TTC             4075                                                                           - (2) INFORMATION FOR SEQ ID NO:68:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 4909 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68:                                 - GAGCTCGCGG CCGCCTATCA AAAGTCTTAA TGAGTTAGGT GTAGATAGTA TA - #GATATTAC          60                                                                           - TACAAAGGTA TTCATATTTC CTATCAATTC TAAAGTAGAT GATATTAATA AC - #TCAAAGAT         120                                                                           - GATGATAGTA GATAATAGAT ACGCTCATAT AATGACTGCA AATTTGGACG GT - #TCACATTT         180                                                                           - TAATCATCAC GCGTTCATAA GTTTCAACTG CATAGATCAA AATCTCACTA AA - #AAGATAGC         240                                                                           - CGATGTATTT GAGAGAGATT GGACATCTAA CTACGCTAAA GAAATTACAG TT - #ATAAATAA         300                                                                           - TACATAATGG ATTTTGTTAT CATCAGTTAT ATTTAACATA AGTACAATAA AA - #AGTATTAA         360                                                                           - ATAAAAATAC TTACTTACGA AAAAATGACT AATTAGCTAT AAAAACCCGG GG - #GATCCTTA         420                                                                           - ATTAATTAGT TATTAGACAA GGTGAAAACG AAACTATTTG TAGCTTAATT AA - #TTAGCTGC         480                                                                           - AGGGCTGCAG GAATTCTAGC AATAAAAACT ATTCCTCCGT GTTCTTAATC TT - #CTCGATCT         540                                                                           - TTTGGAGGAT GTTCTGCACG GCGTCCGACG GCGTTTTGGC GCCCCCCATG CC - #GGCAGAAC         600                                                                           - CCGGTTGCGG CCCCGTACCG CTCTTCTGGG GCGACGATAG GTCGAAAGCC AC - #CGTTTTCA         660                                                                           - TGCCCGTCGT GCTCTTGACG GGGGAACCTA CGGCGGCGGT CCCCGTCGAG CG - #GCGTGATT         720                                                                           - GCAAAGCCGC GCTCGCCCCC GGTTTCAGGA TGGAGGGGGA GGCCACAGGC GG - #CGCATTCG         780                                                                           - ATACGCTGCT TTTGGCCGTA GACGACGGTG GGTAAACGGT GGTTACCGCG GG - #ATACGTCG         840                                                                           - GCGTGGTCGA GGCGGCCCGG CTGGTGCCGG ACAGGCGACC CGGCGCGCTA CC - #GCTCACGG         900                                                                           - GTACCGAGGG CGGTCGACCT ACCACCGCCT TGCCGCCCAA AGTAGGTTTC AA - #AGAAGGAA         960                                                                           - CACCGACGCG GCTGCCCCGA CCTTTCACCG GAGACGGAGG GGCACTCTTG GC - #CGGGGACG        1020                                                                           - GAGAGGCTGA CGAAAGCATG GACAGCGGCG ACGTGACGGG GGACACGACA TC - #ATCCTCCG        1080                                                                           - TGGGCGACAA AACGGACGCC GAGGCTGACG GCTGTCGAGC CGAAGCGGAA GA - #GGTTCTCG        1140                                                                           - CGCCAGAAGT CACGTTCCTT GATGACGTTG TTTTAGACGA AGCCGGTTGA GG - #TTGCAACA        1200                                                                           - GCGTGGCGGG TACCGTCGAC GGCGTGCCCG ATACCTGTTT CTCTACCCTT CC - #CTGAACCG        1260                                                                           - GTGTCGACGT CACCGTCTGC GCTCGGGCGG ACGCGTGCGG CGTCGCGACT CG - #CTTGCCCA        1320                                                                           - GCACCGGTTT CTGGCTCGTG GATGTCGTCG TCATTGGAGA CGATAACTTA GC - #TTTACGTA        1380                                                                           - TTCTGGACGG CGTCGACTGC TCGGGCGTCT GACTGGGAGG CGAAATGACG TC - #GTTGTAAT        1440                                                                           - CGGACGACGG TGTTGTGTGT CCCAGGCTGA CGACGGAGCC GGTGTCCGAG GA - #GTCGTCGT        1500                                                                           - CTTCCTCCTC GCTGTCTTCG ACCGGTGACT CTGCAGTTTG GTCCCTTAAA GC - #CCAAACCT        1560                                                                           - CATCAGCGGC GTTCTGAGAC GCTGTTTGTG TCACCGCGGC GCGTGGAGTC GA - #CGGCCTCC        1620                                                                           - GAGGGGTGGT GGACACGTTG TTTTGAGAAG TCGTGGAAGT CGTAGGCATC CT - #GAAGGGAT        1680                                                                           - TGTAAGCCAG GTGAGGATTC TTGAGGGCCC ACGCGCGTTC GCGCGGCCAG TT - #GGCGGGGT        1740                                                                           - TCATATCCCC GGGCAACGGC GCCGTCGGAG CCCAGGGCGA GTTACCGTTG AC - #CGGGGTTT        1800                                                                           - GGGTACCCGC GAAGGTAGGT GTCGGGGCCG GAGCGGGGGC CGTGGAAGGA TT - #GACAGGCG        1860                                                                           - TCGGCGTGAG GATGGCAGCG CCGGCGCCAG CAGGGACGTT AACTCCGGCG CC - #GAACGTCA        1920                                                                           - ACGTCGGTTG CTCGAACTTG TACGCGGTGG TGACGGGCGG TTTGGCGCTC GT - #CTCGGTAT        1980                                                                           - CCGTGATGTC CACCAGCGTG TCGGTGAAAC GCGGATCTTG ACGGTTGGGG GG - #ATAGCCAT        2040                                                                           - CCGAGCTGTC GGAATCCTCG TCGCCCGAGA AAAGATCCCC TCTTGTCTCC GT - #GAGCGGCC        2100                                                                           - TCACGTCCCA CGCGCTGTCC CGACGGACCC TTCCCGGGCT GGCCTTGGTT AC - #CTGCGGGG        2160                                                                           - AGACGAGACT GAAAGCCGCG TGACGCTGTT GTTGCTGCGG GATGTTCAAG GG - #ACCGCTGG        2220                                                                           - TCGGTTTCTG ACTGCCCGAG GATAACATGC CGCTGAAAAT GCTGGAAACA CC - #GTTGCCAC        2280                                                                           - TAGCGGCGCC CTTGCCGCTA GTTCCCGGTT TCTTGATGGG CGTAAAGATG TT - #TTTCTCGT        2340                                                                           - CATCATCATC GTCGTCGTCC TCATCGGCAC TGGAGCCAAA GAGCCTCCGG GA - #GGCGCCCG        2400                                                                           - GTTTACGTGT CGGGGGCGGC GGTTGCTGCT GACGTTGCTG CAGGTTCTGC TG - #CCTCTCCT        2460                                                                           - CCCAAGCCTT CAGCTGCTGT TTCTCACGCT GCACCACCTC GTCGTCCACC CG - #TTTCTGCC        2520                                                                           - GCTCGCGACG CTTTTCCTCT TCGTCGTAAT AGCCGACGCG CGCCGAACGG GC - #GGCGTGGG        2580                                                                           - CGTCGGCGGC CGGTGCCAGA GAACCATGGG CCTCGAAGCG GAACGGTTTG TG - #TCCCTTCC        2640                                                                           - AGGGACTGGC GATCCAGCTC CAGCCGTCCA GCGGCTGCGT GGGGACATGT TT - #CTTGGGTA        2700                                                                           - CCGACGAGAA GGCTGAACCG CCGCCGAGCG AGAGGAGATT GGCGTCATCG TC - #AAACTCCA        2760                                                                           - ACGACGGCGG GCGCGCGCCC AAAAAGGTGT GCGCCGACTG CGGGAAGCTG TC - #CACGTAGA        2820                                                                           - TGTCAAAGTC CTCGATGAGC AGCTCCAGCA GCGTGTCGGC CGAGTCACCG TT - #TTCCACGG        2880                                                                           - CGTGTTTGAG GATATTGCGA CAGTAGTTGG AATCAAAGGA AAGGCACATG CG - #CAGCTCCT        2940                                                                           - TGACCAGCAG CTTGCAGCGC TCCTGAATGC GCGCCAGACA TTTGCGCTCC AG - #CTCCTCCC        3000                                                                           - AAGACCTGCG CACGTTCATG ATGAGACGGC CCGTGTACAC GAGCTTGTTG AC - #GGCGTTGA        3060                                                                           - CCAGCGCCGT GTTGGCGTGC CGGTCCAGGT TAAGGTCGAG CGGTTTCACG CA - #GAACATGT        3120                                                                           - TACGGCGCAC ACCCTCCAGG TTTTCTTCAA TGCGCTGCAC CTCCGTATCC TT - #GAGGTGCA        3180                                                                           - CAAAAGCGAT GGGTTCCGTC TGGCCGATGG CTGTGACCAG CGTCTCGCGC AC - #CGACATCT        3240                                                                           - TGGCCAGAAT GACCGCGCTT ACGAGCGCGC GCTCCACAAT CTCAGCATCG TG - #GCGTACGT        3300                                                                           - CCGTATCGAA TTCGGTACGG TCTAGCACAG CCAGGTGGTC ACGCGCCTTA CC - #ACGATCAC        3360                                                                           - CGAACGGGTA AGTGTAGCCG CGACGCGCCA CGGCCGCGCA ACGCACCTCG AA - #CTCCTCGA        3420                                                                           - GAACCGAGGA GAGGTCGGGG TTGTGGAAAC GCAGCTCGCG GTAGTATCCC AA - #CCAAAGCA        3480                                                                           - TGAGCTCGTT GAACAGCACC GTACGCCGGT GCAGGCGTTT TTCGCCACAT TT - #TTTCAGGA        3540                                                                           - TCTTGGGGTG TGCCTCGAGA TCCACGTCGG GCTTTTGCGT GAGATGGCGC AG - #AAAGTTGA        3600                                                                           - CCAGGGCCAC CACATCGCGC CGCTGTAGAC CGATAAACTG CAAACTCATT TT - #ATATTGTA        3660                                                                           - ATTATATATT TTCAATTTTG AAATCCCAAA ATATTATCAT ATCTTCCCAA TA - #AAGCTAGA        3720                                                                           - TTCTTTTTAT TGATTAACTA GTCAAATGAG TATATATAAT TGAAAAAGTA AA - #ATATAAAT        3780                                                                           - CATATAATAA TGAAACGAAA TATCAGTAAT AGACAGGAAC TGGCAGATTC TT - #CTTCTAAT        3840                                                                           - GAAGTAAGTA CTGCTAAATC TCCAAAATTA GATAAAAATG ATACAGCAAA TA - #CAGCTTCA        3900                                                                           - TTCAACGAAT TACCTTTTAA TTTTTTCAGA CACACCTTAT TACAAACTAA CT - #AAGTCAGA        3960                                                                           - TGATGAGAAA GTAAATATAA ATTTAACTTA TGGGTATAAT ATAATAAAGA TT - #CATGATAT        4020                                                                           - TAATAATTTA CTTAACGATG TTAATAGACT TATTCCATCA ACCCCTTCAA AC - #CTTTCTGG        4080                                                                           - ATATTATAAA ATACCAGTTA ATGATATTAA AATAGATTGT TTAAGAGATG TA - #AATAATTA        4140                                                                           - TTTGGAGGTA AAGGATATAA AATTAGTCTA TCTTTCACAT GGAAATGAAT TA - #CCTAATAT        4200                                                                           - TAATAATTAT GATAGGAATT TTTTAGGATT TACAGCTGTT ATATGTATCA AC - #AATACAGG        4260                                                                           - CAGATCTATG GTTATGGTAA AACACTGTAA CGGGAAGCAG CATTCTATGG TA - #ACTGGCCT        4320                                                                           - ATGTTTAATA GCCAGATCAT TTTACTCTAT AAACATTTTA CCACAAATAA TA - #GGATCCTC        4380                                                                           - TAGATATTTA ATATTATATC TAACAACAAC AAAAAAATTT AACGATGTAT GG - #CCAGAAGT        4440                                                                           - ATTTTCTACT AATAAAGATA AAGATAGTCT ATCTTATCTA CAAGATATGA AA - #GAAGATAA        4500                                                                           - TCATTTAGTA GTAGCTACTA ATATGGAAAG AAATGTATAC AAAAACGTGG AA - #GCTTTTAT        4560                                                                           - ATTAAATAGC ATATTACTAG AAGATTTAAA ATCTAGACTT AGTATAACAA AA - #CAGTTAAA        4620                                                                           - TGCCAATATC GATTCTATAT TTCATCATAA CAGTAGTACA TTAATCAGTG AT - #ATACTGAA        4680                                                                           - ACGATCTACA GACTCAACTA TGCAAGGAAT AAGCAATATG CCAATTATGT CT - #AATATTTT        4740                                                                           - AACTTTAGAA CTAAAACGTT CTACCAATAC TAAAAATAGG ATACGTGATA GG - #CTGTTAAA        4800                                                                           - AGCTGCAATA AATAGTAAGG ATGTAGAAGA AATACTTTGT TCTATACCTT CG - #GAGGAAAG        4860                                                                           #             4909TAAGT TTAATCAAAC TTGTATTTAT GAAGGTACC                        - (2) INFORMATION FOR SEQ ID NO:69:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 3567 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69:                                 - AAGACTAATT TGTAAACCAT CTTACTCAAA ATATGTAACA ATAGTACGAT GC - #AATGAGTA          60                                                                           - AGACAATAGG AAATCTATCT TATATACACA TAATTATTCT ATCAATTTTA CC - #AATTAGTT         120                                                                           - AGTGTAATGT TATAAAAACT AATTAATCAC TCGAGCCCCT AGCAATAAAA AC - #TATTCCTC         180                                                                           - CGTGTTCTTA ATCTTCTCGA TCTTTTGGAG GATGTTCTGC ACGGCGTCCG AC - #GGCGTTTT         240                                                                           - GGCGCCCCCC ATGCCGGCAG AACCCGGTTG CGGCCCCGTA CCGCTCTTCT GG - #GGCGACGA         300                                                                           - TAGGTCGAAA GCCACCGTTT TCATGCCCGT CGTGCTCTTG ACGGGGGAAC CT - #ACGGCGGC         360                                                                           - GGTCCCCGTC GAGCGGCGTG ATTGCAAAGC CGCGCTCGCC CCCGGTTTCA GG - #ATGGAGGG         420                                                                           - GGAGGCCACA GGCGGCGCAT TCGATACGCT GCTTTTGGCC GTAGACGACG GT - #GGGTAAAC         480                                                                           - GGTGGTTACC GCGGGATACG TCGGCGTGGT CGAGGCGGCC CGGCTGGTGC CG - #GACAGGCG         540                                                                           - ACCCGGCGCG CTACCGCTCA CGGGTACCGA GGGCGGTCGA CCTACCACCG CC - #TTGCCGCC         600                                                                           - CAAAGTAGGT TTCAAAGAAG GAACACCGAC GCGGCTGCCC CGACCTTTCA CC - #GGAGACGG         660                                                                           - AGGGGCACTC TTGGCCGGGG ACGGAGAGGC TGACGAAAGC ATGGACAGCG GC - #GACGTGAC         720                                                                           - GGGGGACACG ACATCATCCT CCGTGGGCGA CAAAACGGAC GCCGAGGCTG AC - #GGCTGTCG         780                                                                           - AGCCGAAGCG GAAGAGGTTC TTGCGCCAGA AGTCACGTTC CTTGATGACG TT - #GTTTTAGA         840                                                                           - CGAAGCCGGT TGAGGTTGCA ACAGCGTGGC GGGTACCGTC GACGGCGTGC CC - #GATACCTG         900                                                                           - TTTCTCTACC CTTCCCTGAA CCGGTGTCGA CGTCACCGTC TGCGCTCGGG CG - #GACGCGTG         960                                                                           - CGGCGTCGCG ACTCGCTTGC CCAGCACCGG TTTCTGGCTC GTGGATGTCG TC - #GTCATTGG        1020                                                                           - AGACGATAAC TTAGCTTTAC GTATTCTGGA CGGCGTCGAC TGCTCGGGCG TC - #TGACTGGG        1080                                                                           - AGGCGAAATG ACGTCGTTGT AATCGGACGA CGGTGTTGTG TGTCCCAGGC TG - #ACGACGGA        1140                                                                           - GCCGGTGTCC GAGGAGTCGT CGTCTTCCTC CTCGCTGTCT TCGACCGGTG AC - #TCTGCAGT        1200                                                                           - TTGGTCCCTT AAAGCCCAAA CCTCATCAGC GGCGTTCTGA GACGCTGTTT GT - #GTCACCGC        1260                                                                           - GGCGCGTGGA GTCGACGGCC TCCGAGGGGT GGTGGACACG TTGTTTTGAG AA - #GTCGTGGA        1320                                                                           - AGTCGTAGGC ATCCTGAAGG GATTGTAAGC CAGGTGAGGA TTCTTGAGGG CC - #CACGCGCG        1380                                                                           - TTCGCGCGGC CAGTTGGCGG GGTTCATATC CCCGGGCAAC GGCGCCGTCG GA - #GCCCAGGG        1440                                                                           - CGAGTTACCG TTGACCGGGG TTTGGGTACC CGCGAAGGTA GGTGTCGGGG CC - #GGAGCGGG        1500                                                                           - GGCCGTGGAA GGATTGACAG GCGTCGGCGT GAGGATGGCA GCGCCGGCGC CA - #GCAGGGAC        1560                                                                           - GTTAACTCCG GCGCCGAACG TCAACGTCGG TTGCTCGAAC TTGTACGCGG TG - #GTGACGGG        1620                                                                           - CGGTTTGGCG CTCGTCTCGG TATCCGTGAT GTCCACCAGC GTGTCGGTGA AA - #CGCGGATC        1680                                                                           - TTGACGGTTG GGGGGATAGC CATCCGAGCT GTCGGAATCC TCGTCGCCCG AG - #AAAAGATC        1740                                                                           - CCCTCTTGTC TCCGTGAGCG GCCTCACGTC CCACGCGCTG TCCCGACGGA CC - #CTTCCCGG        1800                                                                           - GCTGGCCTTG GTTACCTGCG GGGAGACGAG ACTGAAAGCC GCGTGACGCT GT - #TGTTGCTG        1860                                                                           - CGGGATGTTC AAGGGACCGC TGGTCGGTTT CTGACTGCCC GAGGATAACA TG - #CCGCTGAA        1920                                                                           - AATGCTGGAA ACACCGTTGC CACTAGCGGC GCCCTTGCCG CTAGTTCCCG GT - #TTCTTGAT        1980                                                                           - GGGCGTAAAG ATGTTTTTCT CGTCATCATC ATCGTCGTCG TCCTCATCGG CA - #CTGGAGCC        2040                                                                           - AAAGAGCCTC CGGGAGGCGC CCGGTTTACG TGTCGGGGGC GGCGGTTGCT GC - #TGACGTTG        2100                                                                           - CTGCAGGTTC TGCTGCCTCT CCTCCCAAGC CTTCAGCTGC TGTTTCTCAC GC - #TGCACCAC        2160                                                                           - CTCGTCGTCC ACCCGTTTCT GCCGCTCGCG ACGCTTTTCC TCTTCGTCGT AA - #TAGCCGAC        2220                                                                           - GCGCGCCGAA CGGGCGGCGT GGGCGTCGGC GGCCGGTGCC AGAGAACCAT GG - #GCCTCGAA        2280                                                                           - GCGGAACGGT TTGTGTCCCT TCCAGGGACT GGCGATCCAG CTCCAGCCGT CC - #AGCGGCTG        2340                                                                           - CGTGGGGACA TGTTTCTTGG GTACCGACGA GAAGGCTGAA CCGCCGCCGA GC - #GAGAGGAG        2400                                                                           - ATTGGCGTCA TCGTCAAACT CCAACGACGG CGGGCGCGCG CCCAAAAAGG TG - #TGCGCCGA        2460                                                                           - CTGCGGGAAG CTGTCCACGT AGATGTCAAA GTCCTCGATG AGCAGCTCCA GC - #AGCGTGTC        2520                                                                           - GGCCGAGTCA CCGTTTTCCA CGGCGTGTTT GAGGATATTG CGACAGTAGT TG - #GAATCAAA        2580                                                                           - GGAAAGGCAC ATGCGCAGCT CCTTGACCAG CAGCTTGCAG CGCTCCTGAA TG - #CGCGCCAG        2640                                                                           - ACATTTGCGC TCCAGCTCCT CCCAAGACCT GCGCACGTTC ATGATGAGAC GG - #CCCGTGTA        2700                                                                           - CACGAGCTTG TTGACGGCGT TGACCAGCGC CGTGTTGGCG TGCCGGTCCA GG - #TTAAGGTC        2760                                                                           - GAGCGGTTTC ACGCAGAACA TGTTACGGCG CACACCCTCC AGGTTTTCTT CA - #ATGCGCTG        2820                                                                           - CACCTCCGTA TCCTTGAGGT GCACAAAAGC GATGGGTTCC GTCTGGCCGA TG - #GCTGTGAC        2880                                                                           - CAGCGTCTCG CGCACCGACA TCTTGGCCAG AATGACCGCG CTTACGAGCG CG - #CGCTCCAC        2940                                                                           - AATCTCAGCA TCGTGGCGTA CGTCCGTATC GAATTCGGTA CGGTCTAGCA CA - #GCCAGGTG        3000                                                                           - GTCACGCGCC TTACCACGAT CACCGAACGG GTAAGTGTAG CCGCGACGCG CC - #ACGGCCGC        3060                                                                           - GCAACGCACC TCGAACTCCT CGAGAACCGA GGAGAGGTCG GGGTTGTGGA AA - #CGCAGCTC        3120                                                                           - GCGGTAGTAT CCCAACCAAA GCATGAGCTC GTTGAACAGC ACCGTACGCC GG - #TGCAGGCG        3180                                                                           - TTTTTCGCCA CATTTTTTCA GGATCTTGGG GTGTGCCTCG AGATCCACGT CG - #GGCTTTTG        3240                                                                           - CGTGAGATGG CGCAGAAAGT TGACCAGGGC CACCACATCG CGCCGCTGTA GA - #CCGATAAA        3300                                                                           - CTGCAAACTC ATTTTATATT GTAATTATAT ATTTTCAATT TTGAAATCCC AA - #AATATTAT        3360                                                                           - CATATCTTCC CAATAAAGCT AGGGGGAATT CGGATCCTCG CGACTGCAGG GT - #ACCTGAGT        3420                                                                           - AGCTAATTTT TAAACAAAAA TGTGGGAGAA TCTAATTAGT TTTTCTTTAC AC - #AATTGACG        3480                                                                           - TACATGAGTC TGAGTTCCTT GTTTTTGCTA ATTATTTCAT CCAATTTATT AT - #TCTTGACG        3540                                                                           #           3567   TATA GGAGTCA                                                - (2) INFORMATION FOR SEQ ID NO:70:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 4893 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70:                                 - CTGCAGGTCG ACGGATCTGA GAATGGATGA TTCTCCAGCC GAAACATATT CT - #ACCATGGC          60                                                                           - TCCGTTTAAT TTGTTGATGA AGATGGATTC ATCCTTAAAT GTTTTCTCTG TA - #ATAGTTTC         120                                                                           - CACCGAAAGA CTATGCAAAG AATTTGGAAT GCGTTCCTTG TGCTTAATGT TT - #CCATAGAC         180                                                                           - GGCTTCTAGA AGTTGATACA ACATAGGACT AGCCGCGGTA ACTTTTATTT TT - #AGAAAGTA         240                                                                           - TCCATCGCTT CTATCTTGTT TAGATTTATT TTTATAAAGT TTAGTCTCTC CT - #TCCAACAT         300                                                                           - AATAAAAGTG GAAGTCATTT GACTAGATAA ACTATCAGTA AGTTTTATAG AG - #ATAGACGA         360                                                                           - ACAATTAGCG TATTGAGAAG CATTTAGTGT AACGTATTCG ATACATTTTG CA - #TTAGATTT         420                                                                           - ACTAATCGAT TTTGCATACT CTATAACACC CGCACAAGTC TGTAGAGAAT CG - #CTAGATGC         480                                                                           - AGTAGGTCTT GGTGAAGTTT CAACTCTCTT CTTGATTACC TTACTCATGA TT - #AAACCTAA         540                                                                           - ATAATTGTAC TTTGTAATAT AATGATATAT ATTTTCACTT TATCTCATTT GA - #GAATAAAA         600                                                                           - AGATCACAAA AATTAACTAA TCAGGATCTC GAGATAAAAA TCAGCATGTC TT - #GAGCATGC         660                                                                           - GGTAGAGCAG ATAGATGCCG ATGATGGCCG ATAGCGCGTA GACGGACATC AT - #GAGGAGAC         720                                                                           - GACTGTCGGT AGCGTCCACG ACGACGTCAG TTACTTCTAG GACCGTACCG TT - #TTTCAAAA         780                                                                           - GCATGAGGTA GTGAGTTCGC GGAGATGAGA CCACCACTTC GTTGTAGGGA TC - #CAGGGCGA         840                                                                           - AAAGGACGTC GTCCGAGTCG TGCATGTACA TGATGTTGAT GACGCCTTGC GT - #GTCGTCGT         900                                                                           - ATTCTAGTAG GGCGCTTTGG CAAAAGGCGC AGTTTTCTAG GGAAATGTTG AG - #CGCCGCTG         960                                                                           - TGATGCTGTG TGTGGTATGC ATGTTGCGCG TCAGTTCGCA TTTAGTTTGA CT - #GTCCGTCT        1020                                                                           - GGGTGATGAT GAGGCTCTGG CCTACGACGG TGGTGGAGAC AGGGTAGGAG AT - #ACCTTTGA        1080                                                                           - TCAGGTACTG GTTTGTTACG ACATAACTGA CGTGTTCGGA GACGGTCAGC GC - #GGAGAAGG        1140                                                                           - ATTCGCCGAG CGGCAGACAA AACAGGTCGG GGAAGGTTTC TAGCGTGCTT GG - #TTGCATGG        1200                                                                           - TAGATAGGAT GGAGAGGGCG GCGGGAACGG TAGTGGGGAC GGTGGCATCG GG - #GAAGAGAC        1260                                                                           - GTGTGAGGCG TTCGAGCGAG TGATCGCGTC GCCCGCTACT GGAACAGGGT GT - #GTACAGGT        1320                                                                           - CGCTGAGGTA TTCGTGGTGC GGATGAGCTA GCAACTGCGT AAAGTGTGAT AG - #CTCGGCTA        1380                                                                           - ATGAACAGAG GCCCGTTTCT ACGATGAAGA TTTCGCGTCT CTCCGTCGTA TG - #TACTAGCA        1440                                                                           - TGGAGTGGAC GAGGCTGCCC ATGAGGTAGA GTTCTTGACG CGCGAAGGCT GA - #AAGAAAAG        1500                                                                           - AGGCCAGGTG CGTTTTGTGT AGTTTTAGGG CAAAGTCGGC GATCTGTCGT AG - #TGCCCACT        1560                                                                           - GGGGGATGAG ATGTTGCTGA TTCTGTTTAG AGAGTATGTA GACCAGGCGT AC - #GAGGCTGG        1620                                                                           - TGATGTCGGT GATCTGATTC GGTGTCCAAA GGGCTCGTTT GGCCAGGTCC AC - #GGCCGTGG        1680                                                                           - GATACAGCAG CAACGTGGTG CGTGGTGGTG TTTGTGAGAG GCAGGTGATC AT - #AAATTCTT        1740                                                                           - GTATTTGTAA GAGTGCGGCC TGGCGGTCTA GGGCCCGTGG GACGGAGACT TG - #GGCGCCGG        1800                                                                           - CCTCTTCTTG TCGGGCTGCT GCGAACAGTG CTAATGCGTA GGCGAAGGCC AT - #TTCTACCG        1860                                                                           - TGCGGCGGTC CAGCATCTGA CATCGACCGC TTTTGAGTAC ATCCACGGCG TA - #ACGGTGAA        1920                                                                           - AGCTGTTACG TAGTAGTGCG CTGAGGTCCA GGTAGTTGAA GTCAAGTGCG GC - #GTCAAGAA        1980                                                                           - AGTCCGGGTC TTTGAGATAA GAGTGACGGT TCAGTTGATC TTTCTTAACT AG - #CACCAGGA        2040                                                                           - GCTCGTGTTT TTCAGTTTGT CGTAGTATAA AGTTGTCGCG TTGATAGGGC GC - #TTTAAAGA        2100                                                                           - GTACGCGTGG AAGATGGCCG AAGATAAGCA GCATGGGTGT GTCGTCGTCT AT - #GGACACCG        2160                                                                           - TAACTACGAA GAAGTCCTCG GTCAGTGTTA TTTTAACGTA ACGTAGTTCG TC - #GATGAGGT        2220                                                                           - AAAAGCCTTG GTGCAAACAA GGTGTGACGG TGCTGAATAG TAGATCGTGT CC - #ATCAAAGA        2280                                                                           - GGATACAGGT CTGGTTAAAG TGTGGTCGGT GTAGTCCTGA GGTGGTATGT GA - #TTCTGTCC        2340                                                                           - AGCCGTGTGG AGTGGTTTGC GGTGGCATCC AAACGTGAGG TATTGACAGG TC - #AATGGGTG        2400                                                                           - GTGGCACAGT GGTGGGCTGT TCACCTAGGC TGTCCTGTGC CTTTAGCTGC TG - #CGAAAAAG        2460                                                                           - ATCGGTAGCT GGCCAGGTCT TTGGATACCA GCGCGTAAGT GTTAAGTCTC TG - #TTGGTATC        2520                                                                           - TTTCCAGGGT TTCGGTCAGA TCTACCTGGT TCAGAAACTG CTCCGCCAGA GG - #ACCCGCAA        2580                                                                           - AAAGACATCG AGGCATATGG AATACATAGT ATTGATTATA GCTTTGGAAA AA - #GTTGAAAC        2640                                                                           - TGATGGCGTT TTCCCTGACG ACCGTGCTGT TACGGAGGCT GCTATTGTAG GT - #ACACTGGG        2700                                                                           - TGGTGTTTTC ACGCAGGAAG CGGATGGGTC TCCCGTAGGT GTTGAGCAGT AG - #GTGAAACG        2760                                                                           - CTTTGTCCAG CGGTTCGGAT ATGGCTTCTG CGCCATATCG TGACGAAAGT AG - #GTGGCTGA        2820                                                                           - GGAGACAGAC GGCGAGGACG ATGAGGTAGG AGGGGAGCCC GGGCCGCATT TT - #ATATTGTA        2880                                                                           - ATTATATATT TTCAATTTTG AAATCCCAAA ATATTATCAT ATTCTTCCCA AT - #AAACTCGA        2940                                                                           - GATCCTTCTT TATTCTATAC TTAAAAAGTG AAAATAAATA CAAAGGTTCT TG - #AGGGTTGT        3000                                                                           - GTTAAATTGA AAGCGAGAAA TAATCATAAA TTATTTCATT ATCGCGATAT CC - #GTTAAGTT        3060                                                                           - TGTATCGTAA TGAAACAGAT TAAGGTTCGA GTGGACATGG TGCGGCATAG AA - #TCAAGGAG        3120                                                                           - CACATGCTGA AAAAATATAC CCAGACGGAA GAGAAATTCA CTGGCGCCTT TA - #ATATGATG        3180                                                                           - GGAGGATGTT TGCAGAATGC CTTAGATATC TTAGATAAGG TTCATGAGCC TT - #TCGAGGAG        3240                                                                           - ATGAAGTGTA TTGGGCTAAC TATGCAGAGC ATGTATGAGA ACTACATTGT AC - #CTGAGGAT        3300                                                                           - AAGCGGGAGA TGTGGATGGC TTGTATTAAG GAGCTGCATG ATGTGAGCAA GG - #GCGCCGCT        3360                                                                           - AACAAGTTGG GGGGTGCACT GCAGGCTAAG GCCCGTGCTA AAAAGGATGA AC - #TTAGGAGA        3420                                                                           - AAGATGATGT ATATGTGCTA CAGGAATATA GAGTTCTTTA CCAAGAACTC AG - #CCTTCCCT        3480                                                                           - AAGACCACCA ATGGCTGCAG TCAGGCCATG GCGGCACTGC AGAACTTGCC TC - #AGTGCTCC        3540                                                                           - CCTGATGAGA TTATGGCTTA TGCCCAGAAA ATATTTAAGA TTTTGGATGA GG - #AGAGAGAC        3600                                                                           - AAGGTGCTCA CGCACATTGA TCACATATTT ATGGATATCC TCACTACATG TG - #TGGAAACA        3660                                                                           - ATGTGTAATG AGTACAAGGT CACTAGTGAC GCTTGTATGA TGACCATGTA CG - #GGGGCATC        3720                                                                           - TCTCTCTTAA GTGAGTTCTG TCGGGTGCTG TGCTGCTATG TCTTAGAGGA GA - #CTAGTGTG        3780                                                                           - ATGCTGGCCA AGCGGCCTCT GATAACCAAG CCTGAGGTTA TCAGTGTAAT GA - #AGCGCCGC        3840                                                                           - ATTGAGGAGA TCTGCATGAA GGTCTTTGCC CAGTACATTC TGGGGGCCGA TC - #CTCTGAGA        3900                                                                           - GTCTGCTCTC CTAGTGTGGA TGACCTACGG GCCATCGCCG AGGAGTCAGA TG - #AGGAAGAG        3960                                                                           - GCTATTGTAG CCTACACTTT GGCCACCGCT GGTGTCAGCT CCTCTGATTC TC - #TGGTGTCA        4020                                                                           - CCCCCAGAGT CCCCTGTACC CGCGACTATC CCTCTGTCCT CAGTAATTGT GG - #CTGAGAAC        4080                                                                           - AGTGATCAGG AAGAAAGTGA GCAGAGTGAT GAGGAAGAGG AGGAGGGTGC TC - #AGGAGGAG        4140                                                                           - CGGGAGGACA CTGTGTCTGT CAAGTCTGAG CCAGTGTCTG AGATAGAGGA AG - #TTGCCCCA        4200                                                                           - GAGGAAGAGG AGGATGGTGC TGAGGAACCC ACCGCCTCTG GAGGTAAGAG TA - #CCCACCCT        4260                                                                           - ATGGTGACTA GAAGCAAGGC TGACCAGTAA TTTTTATCTC GAGCCCGGGA GA - #TCTTAGCT        4320                                                                           - AACTGATTTT TCTGGGAAAA AAATTATTTA ACTTTTCATT AATAGGGATT TG - #ACGTATGT        4380                                                                           - AGCGTACAAA ATTATCGTTC CTGGTATATA GATAAAGAGT CCTATATATT TG - #AAAATCGT        4440                                                                           - TACGGCTCGA TTAAACTTTA ATGATTGCAT AGTGAATATA TCATTAGGAT TT - #AACTCCTT        4500                                                                           - GACTATCATG GCGGCGCCAG AAATTACCAT CAAAAGCATT AATACAGTTA TG - #CCGATCGC        4560                                                                           - AGTTAGAACG GTTATAGCAT CCACCATTTA TATCTAAAAA TTAGATCAAA GA - #ATATGTGA        4620                                                                           - CAAAGTCCTA GTTGTATACT GAGAATTGAC GAAACAATGT TTCTTACATA TT - #TTTTTCTT        4680                                                                           - ATTAGTAACT GACTTAATAG TAGGAACTGG AAAGCTAGAC TTGATTATTC TA - #TAAGTATA        4740                                                                           - GATACCCTTC CAGATAATGT TCTCTTTGAT AAAAGTTCCA GAAAATGTAG AA - #TTTTTTAA        4800                                                                           - AAAGTTATCT TTTGCTATTA CCAAGATTGT GTTTAGACGC TTATTATTAA TA - #TGAGTAAT        4860                                                                           #       4893       CTAG ATATGGGGAA TTC                                         - (2) INFORMATION FOR SEQ ID NO:71:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 6749 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:71:                                 - GAGCTCGCGG CCGCCTATCA AAAGTCTTAA TGAGTTAGGT GTAGATAGTA TA - #GATATTAC          60                                                                           - TACAAAGGTA TTCATATTTC CTATCAATTC TAAAGTACAT GATATTAATA AC - #TCAAAGAT         120                                                                           - GATGATAGTA GATAATAGAT ACGCTCATAT AATGACTGCA AATTTGGACG GT - #TCACATTT         180                                                                           - TAATCATCAC GCGTTCATAA GTTTCAACTG CATAGATCAA AATCTCACTA AA - #AAGATAGC         240                                                                           - CGATGTATTT GAGAGAGATT GGACATCTAA CTACGCTAAA GAAATTACAG TT - #ATAAATAA         300                                                                           - TACATAATGG ATTTTGTTAT CATCAGTTAT ATTTAACATA AGTACAATAA AA - #AGTATTAA         360                                                                           - ATAAAAATAC TTACTTACGA AAAAATGACT AATTAGCTAT AAAAACCCAA CA - #AAAACTAA         420                                                                           - TCAGCTATCG GGGTTAATTA ATTAGTTATT AGACAAGGTG AAAACGAAAC TA - #TTTGTAGC         480                                                                           - TTAATTAATT AGAGCTTCTT TATTCTATAC TTAAAAAGTG AAAATAAATA CA - #AAGGTTCT         540                                                                           - TGAGGGTTGT GTTAAATTGA AAGCGAGAAA TAATCATAAA TTATTTCATT AT - #GGCGATAT         600                                                                           - CCGTTAAGTT TGTATCGTAA TGGAGTCGCG CGGTCGCCGT TGTCCCGAAA TG - #ATATCCGT         660                                                                           - ACTGGGTCCC ATTTCGGGGC ACGTGCTGAA AGCCGTGTTT AGTCGCGGCG AC - #ACGCCGGT         720                                                                           - GCTGCCGCAC GAGACGCGAC TCCTGCAGAC GGGTATCCAC GTGCGCGTGA GC - #CAGCCCTC         780                                                                           - GCTGATCCTG GTGTCGCAGT ACACGCCCGA CTCGACGCCA TGCCACCGCG GC - #GACAATCA         840                                                                           - GCTGCAGGTG CAGCACACGT ACTTTACGGG CAGCGAGGTG GAGAACGTGT CG - #GTCAACGT         900                                                                           - GCACAACCCC ACGGGCCGGA GCATCTGCCC CAGCCAAGAG CCCATGTCGA TC - #TATGTGTA         960                                                                           - GCGCGCTGCC GCTCAAGATG CTGAACATCC CCAGCATCAA CGTGCACCAC TA - #CCCGTCGG        1020                                                                           - CGGCCGAGCG CAAACACCGA CACCTGCCCG TAGCTGACGC TGTGATTCAC GC - #GTCGGGCA        1080                                                                           - AGCAGATGTG GCAGGCGCGT CTCACGGTCT CGGGACTGGC CTGGACGCGT CA - #GCAGAACC        1140                                                                           - AGTGGAAAGA GCCCGACGTC TACTACACGT CAGCGTTCGT GTTTCCCACC AA - #GGACGTGG        1200                                                                           - CACTGCGGCA CGTGGTGTGC GCGCACGAGC TGGTTTGCTC CATGGAGAAC AC - #GCGCGCAA        1260                                                                           - CCAAGATGCA GGTGATAGGT GACCAGTACG TCAAGGTGTA CCTGGAGTCC TT - #CTGCGAGG        1320                                                                           - ACGTGCCCTC CGGCAAGCTC TTTATGCACG TCACGCTGGG CTCTGACGTG GA - #AGAGGACC        1380                                                                           - TGACGATGAC CCGCAACCCG CAACCCTTCA TGCGCCCCCA CGAGCGCAAC GG - #CTTTACGG        1440                                                                           - TGTTGTGTCC CAAAAATATG ATAATCAAAC CGGGCAAGAT CTCGCACATC AT - #GCTGGATG        1500                                                                           - TGGCTTTTAC CTCACACGAG CATTTTGGGC TGCTGTGTCC CAAGAGCATC CC - #GGGCCTGA        1560                                                                           - GCATCTCAGG TAACCTATTG ATGAACGGGC AGCAGATCTT CCTGGAGGTG CA - #AGCGATAC        1620                                                                           - GCGAGACCGT GGAACTGCGT CAGTACGATC CCGTGGCTGC GCTCTTCTTT TT - #CGATATCG        1680                                                                           - AGCTTGCTGC TGCAGCGCGG GCCTCAGTAC AGCGAACACC CCACCTTCAC CA - #GCCAGTAT        1740                                                                           - CGCATCCAGG GCAAGCTTGA GTACCGACAC ACCTGGGACC GGCACGACGA GG - #GTGCCGCC        1800                                                                           - CAGGGCGACG ACGACGTCTG GACCAGCGGA TCGGACTCCG ACGAGGAACT CG - #TAACCACC        1860                                                                           - GAGGCGCAAG ACGCCCCGCG TTACCGGCGG CGGCGCCATG GCGGGCGCCT CC - #ACTTCCGC        1920                                                                           - GGGCCGCAAA CGCAAATCAG CATCCTCGGC GACGGCGTGC ACGGCGGGCG TT - #ATGACACG        1980                                                                           - CGGCCGCCTT AAGGCCGAGT CCACCGTCGC GCCCGAAGAG GACACCGACG AG - #GATTCCGA        2040                                                                           - CAACGAAATC CACAATCCGG CCGTGTTCAC CTGGCCGCCC TGGCAGGCCG GC - #ATCCTGGC        2100                                                                           - CCGCAACCTG GTGCCCATGG TGGCTACGGT TCAGGGTCAG AATCTGAAGT AC - #CAGGAGTT        2160                                                                           - CTTCTGGGAC GCCAACGACA TCTACCGCAT CTTCGCCGAA TTGGAAGGCG TA - #TGGCAGCC        2220                                                                           - CGCTGCGCAA CCCAAACGTC GCCGCCACCG GCAAGACGCC TTGCCCGGGC CA - #TGCATCGC        2280                                                                           - CTCGACGCCC AAAAAGCACC GAGGTTGATT TTTATGGATC CGGTACCCTC GA - #GGAATTCT        2340                                                                           - AGCAATAAAA ACTATTCCTC CGTGTTCTTA ATCTTCTCGA TCTTTTGGAG GA - #TGTTCTGC        2400                                                                           - ACGGCGTCCG ACGGCGTTTT GGCGCCCCCC ATGCCGGCAG AACCCGGTTG CG - #GCCCCGTA        2460                                                                           - CCGCTCTTCT GGGGCGACGA TAGGTCGAAA GCCACCGTTT TCATGCCCGT CG - #TGCTCTTG        2520                                                                           - ACGGGGGAAC CTACGGCGGC GGTCCCCGTC GAGCGGCGTG ATTGCAAAGC CG - #CGCTCGCC        2580                                                                           - CCCGGTTTCA GGATGGAGGG GGAGGCCACA GGCGGCGCAT TCGATACGCT GC - #TTTTGGCC        2640                                                                           - GTAGACGACG GTGGGTAAAC GGTGGTTACC GCGGGATACG TCGGCGTGGT CG - #AGGCGGCC        2700                                                                           - CGGCTGGTGC CGGACAGGCG ACCCGGCGCG CTACCGCTCA CGGGTACCGA GG - #GCGGTCGA        2760                                                                           - CCTACCACCG CCTTGCCGCC CAAAGTAGGT TTCAAAGAAG GAACACCGAC GC - #GGCTGCCC        2820                                                                           - CGACCTTTCA CCGGAGACGG AGGGGCACTC TTGGCCGGGG ACGGAGAGGC TG - #ACGAAAGC        2880                                                                           - ATGGACAGCG GCGACGTGAC GGGGGACACG ACATCATCCT CCGTGGGCGA CA - #AAACGGAC        2940                                                                           - GCCGAGGCTG ACGGCTGTCG AGCCGAAGCG GAAGAGGTTC TCGCGCCAGA AG - #TCACGTTC        3000                                                                           - CTTGATGACG TTGTTTTAGA CGAAGCCGGT TGAGGTTGCA ACAGCGTGGC GG - #GTACCGTC        3060                                                                           - GACGGCGTGC CCGATACCTG TTTCTCTACC CTTCCCTGAA CCGGTGTCGA CG - #TCACCGTC        3120                                                                           - TGCGCTCGGG CGGACGCGTG CGGCGTCGCG ACTCGCTTGC CCAGCACCGG TT - #TCTGGCTC        3180                                                                           - GTGGATGTCG TCGTCATTGG AGACGATAAC TTAGCTTTAC GTATTCTGGA CG - #GCGTCGAC        3240                                                                           - TGCTCGGGCG TCTGACTGGG AGGCGAAATG ACGTCGTTGT AATCGGACGA CG - #GTGTTGTG        3300                                                                           - TGTCCCAGGC TGACGACGGA GCCGGTGTCC GAGGAGTCGT CGTCTTCCTC CT - #CGCTGTCT        3360                                                                           - TCGACCGGTG ACTCTGCAGT TTGGTCCCTT AAAGCCCAAA CCTCATCAGC GG - #CGTTCTGA        3420                                                                           - GACGCTGTTT GTGTCACCGC GGCGCGTGGA GTCGACGGCC TCCGAGGGGT GG - #TGGACACG        3480                                                                           - TTGTTTTGAG AAGTCGTGGA AGTCGTAGGC ATCCTGAAGG GATTGTAAGC CA - #GGTGAGGA        3540                                                                           - TTCTTGAGGG CCCACGCGCG TTCGCGCGGC CAGTTGGCGG GGTTCATATC CC - #CGGGCAAC        3600                                                                           - GGCGCCGTCG GAGCCCAGGG CGAGTTACCG TTGACCGGGG TTTGGGTACC CG - #CGAAGGTA        3660                                                                           - GGTGTCGGGG CCGGAGCGGG GGCCGTGGAA GGATTGACAG GCGTCGGCGT GA - #GGATGGCA        3720                                                                           - GCGCCGGCGC CAGCAGGGAC GTTAACTCCG GCGCCGAACG TCAACGTCGG TT - #GCTCGAAC        3780                                                                           - TTGTACGCGG TGGTGACGGG CGGTTTGGCG CTCGTCTCGG TATCCGTGAT GT - #CCACCAGC        3840                                                                           - GTGTCGGTGA AACGCGGATC TTGACGGTTG GGGGGATAGC CATCCGAGCT GT - #CGGAATCC        3900                                                                           - TCGTCGCCCG AGAAAAGATC CCCTCTTGTC TCCGTGAGCG GCCTCACGTC CC - #ACGCGCTG        3960                                                                           - TCCCGACGGA CCCTTCCCGG GCTGGCCTTG GTTACCTGCG GGGAGACGAG AC - #TGAAAGCC        4020                                                                           - GCGTGACGCT GTTGTTGCTG CGGGATGTTC AAGGGACCGC TGGTCGGTTT CT - #GACTGCCC        4080                                                                           - GAGGATAACA TGCCGCTGAA AATGCTGGAA ACACCGTTGC CACTAGCGGC GC - #CCTTGCCG        4140                                                                           - CTAGTTCCCG GTTTCTTGAT GGGCGTAAAG ATGTTTTTCT CGTCATCATC AT - #CGTCGTCG        4200                                                                           - TCCTCATCGG CACTGGAGCC AAAGAGCCTC CGGGAGGCGC CCGGTTTACG TG - #TCGGGGGC        4260                                                                           - GGCGGTTGCT GCTGACGTTG CTGCAGGTTC TGCTGCCTCT CCTCCCAAGC CT - #TCAGCTGC        4320                                                                           - TGTTTCTCAC GCTGCACCAC CTCGTCGTCC ACCCGTTTCT GCCGCTCGCG AC - #GCTTTTCC        4380                                                                           - TCTTCGTCGT AATAGCCGAC GCGCGCCGAA CGGGCGGCGT GGGCGTCGGC GG - #CCGGTGCC        4440                                                                           - AGAGAACCAT GGGCCTCGAA GCGGAACGGT TTGTGTCCCT TCCAGGGACT GG - #CGATCCAG        4500                                                                           - CTCCAGCCGT CCAGCGGCTG CGTGGGGACA TGTTTCTTGG GTACCGACGA GA - #AGGCTGAA        4560                                                                           - CCGCCGCCGA GCGAGAGGAG ATTGGCGTCA TCGTCAAACT CCAACGACGG CG - #GGCGCGCG        4620                                                                           - CCCAAAAAGG TGTGCGCCGA CTGCGGGAAG CTGTCCACGT AGATGTCAAA GT - #CCTCGATG        4680                                                                           - AGCAGCTCCA GCAGCGTGTC GGCCGAGTCA CCGTTTTCCA CGGCGTGTTT GA - #GGATATTG        4740                                                                           - CGACAGTAGT TGGAATCAAA GGAAAGGCAC ATGCGCAGCT CCTTGACCAG CA - #GCTTGCAG        4800                                                                           - CGCTCCTGAA TGCGCGCCAG ACATTTGCGC TCCAGCTCCT CCCAAGACCT GC - #GCACGTTC        4860                                                                           - ATGATGAGAC GGCCCGTGTA CACGAGCTTG TTGACGGCGT TGACCAGCGC CG - #TGTTGGCG        4920                                                                           - TGCCGGTCCA GGTTAAGGTC GAGCGGTTTC ACGCAGAACA TGTTACGGCG CA - #CACCCTCC        4980                                                                           - AGGTTTTCTT CAATGCGCTG CACCTCCGTA TCCTTGAGGT GCACAAAAGC GA - #TGGGTTCC        5040                                                                           - GTCTGGCCGA TGGCTGTGAC CAGCGTCTCG CGCACCGACA TCTTGGCCAG AA - #TGACCGCG        5100                                                                           - CTTACGAGCG CGCGCTCCAC AATCTCACCA TCGTGGCGTA CGTCCGTATC GA - #ATTCGGTA        5160                                                                           - CGGTCTAGCA CAGCCAGGTG GTCACGCGCC TTACCACGAT CACCGAACGG GT - #AAGTGTAG        5220                                                                           - CCGCGACGCG CCACGGCCGC GCAACGCACC TCGAACTCCT CGGAACCGAG GA - #GAGGTCGG        5280                                                                           - GGTTGTGGAA ACGCAGCTCG CGGTAGTATC CCAACCAAAG CATGAGCTCG TT - #GAACAGCA        5340                                                                           - CCGTACGCCG GTGCAGGCGT TTTTCGCCAC ATTTTTTCAG GATCTTGGGG TG - #TGCCTCGA        5400                                                                           - GATCCACGTC GGGCTTTTGC GTGAGATGGC GCAGAAAGTT GACCAGGGCC AC - #CACATCGC        5460                                                                           - GCCGCTGTAG ACCGATAAAC TGCAAACTCA TTTTATATTG TAATTATATA TT - #TTCAATTT        5520                                                                           - TGAAATCCCA AAATATTATC ATATCTTCCC AATAAAGCTA GAATTCTTTT TA - #TTGATTAA        5580                                                                           - CTAGTCAAAT GAGTATATAT AATTGAAAAA GTAAAATATA AATCATATAA TA - #ATGAAACG        5640                                                                           - AAATATCAGT AATAGACAGG AACTGGCAGA TTCTTCTTCT AATGAAGTAA GT - #ACTGCTAA        5700                                                                           - ATCTCCAAAA TTAGATAAAA ATGATACACC AAATACAGCT TCATTCAACG AA - #TTACCTTT        5760                                                                           - TAATTTTTTC AGACACACCT TATTACAAAC TAACTAAGTC AGATGATGAG AA - #AGTAAATA        5820                                                                           - TAAATTTAAC TATGGGTATA ATATAATAAA GATTCATGAT ATTAATAATT TA - #CTTAACGA        5880                                                                           - TGTTAATAGA CTTATTCCAT CAACCCCTTC AAACCTTTCT GGATATTATA AA - #ATACCAGT        5940                                                                           - AATGATATTA AAATAGATTG TTTAAGAGAT GTAAATAATT ATTTGGAGGT AA - #AGGATATA        6000                                                                           - AAATTAGTCT ATCTTTCACA TGGAAATGAA TTACCTAATA TTAATAATTA TG - #ATAGGAAT        6060                                                                           - TTTTTAGGAT TTACAGCTGT TATATGTATC AACAATACAG GCAGATCTAT GG - #TTATGGTA        6120                                                                           - AAACACTGTA ACGGGAAGCA GCATTCTATG GTAACTGGCC TATGTTTAAT AG - #CCAGATCA        6180                                                                           - TTTTACTCTA TAAACATTTT ACCACAAATA ATAGGATCCT CTAGATATTT AA - #TATTATAT        6240                                                                           - CTAACAACAA CAAAAAAATT TAACGATGTA TGGCCAGAAG TATTTTCTAC TA - #ATAAAGAT        6300                                                                           - AAAGATAGTC TATCTTATCT ACAAGATATG AAAGAAGATA ATCATTTAGT AG - #TAGCTACT        6360                                                                           - AATATGGAAA GAAATGTATA CAAAAACGTG GAAGCTTTTA TATTAAATAG CA - #TATTACTA        6420                                                                           - GAAGATTTAA AATCTAGACT TAGTATAACA AAACAGTTAA ATGCCAATAT CG - #ATTCTATA        6480                                                                           - TTTCATCATA ACAGTAGTAC ATTAATCAGT GATATACTGA AACGATCTAC AG - #ACTCAACT        6540                                                                           - ATGCAAGGAA TAAGCAATAT GCCAATTATG TCTAATATTT TAACTTTAGA AC - #TAAAACGT        6600                                                                           - TCTACCAATA CTAAAAATAG GATACGTGAT AGGCTGTTAA AAGCTGCAAT AA - #ATAGTAAG        6660                                                                           - GATGTAGAAG AAATACTTTG TTCTATACCT TCGGAGGAAA GAACTTTAGA AC - #AACTTAAG        6720                                                                           #          6749    TTTA TGAAGGTAC                                              - (2) INFORMATION FOR SEQ ID NO:72:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 837 base                                                           (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72:                                 - ATGTGCCGCC GCCCGGATTG CGGCTTCTCT TTCTCACCTG GACCGGTGGC AC - #TGCTGTGG          60                                                                           - TGTTGCCTTC TGCTGCCCAT CGTTTCCTCA GCCACCGTCA GCGTCGCTCC TA - #CCGTCGCC         120                                                                           - GAGAAAGTTC CCGCGGAGTG CCCCGAACTA ACGCGTCGAT GCCTGTTGGG TG - #AGGTGTTT         180                                                                           - CAGGGTGACA AGTATGAAAG TTGGCTGCGC CCGTTGGTGA ATGTTACCAG AC - #GCGATGGC         240                                                                           - CCGCTATCGC AACTTATTCG TTACCGTCCC GTTACGCCGG AGGCCGCCAA CT - #CCGTGCTG         300                                                                           - TTGGACGATG CTTTCCTGGA CACTCTGGCC CTGCTGTACA ACAATCCGGA TC - #AATTGCGG         360                                                                           - GCCTTGCTGA CGCTGTTGAG CTCGGACACA GCGCCGCGCT GGATGACGGT GA - #TGCGCGGT         420                                                                           - TACAGCGAGT GCGGCGATGG CTCGCCGGCC GTGTACACGT GCGTGGACGA CC - #TGTGCCGC         480                                                                           - GGCTACGACC TCACGCGACT GTCATACGGG CGCAGCATCT TCACGGAACA CG - #TGTTAGGC         540                                                                           - TTCGAGCTGG TGCCACCGTC TCTCTTTAAC GTGGTGGTGG CCATACGCAA CG - #AAGCCACG         600                                                                           - CGTACCAACC GCGCCGTGCG TCTGCCCGTG AGCACCGCTG CCGCGCCCGA GG - #GCATCACG         660                                                                           - CTCTTTTACG GCCTGTACAA CGCAGTGAAG GAATTCTGCC TGCGTCACCA GC - #TGGACCCG         720                                                                           - CCGCTGCTAC GCCACCTAGA TAAATACTAC GCCGGACTGC CGCCCGAGCT GA - #AGCAGACG         780                                                                           - CGCGTCAACC TGCCGGCTCA CTCGCGCTAT GGCCCTCAAG CAGTGGATGC TC - #GCTAA            837                                                                           - (2) INFORMATION FOR SEQ ID NO:73:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 5234 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:73:                                 - AAGCTTTTGC GATCAATAAA TGGATCACAA CCAGTATCTC TTAACGATGT TC - #TTCGCAGA          60                                                                           - TGATGATTCA TTTTTTAAGT ATTTGGCTAG TCAAGATGAT GAATCTTCAT TA - #TCTGATAT         120                                                                           - ATTGCAAATC ACTCAATATC TAGACTTTCT GTTATTATTA TTGATCCAAT CA - #AAAAATAA         180                                                                           - ATTAGAAGCC GTGGGTCATT GTTATGAATC TCTTTCAGAG GAATACAGAC AA - #TTGACAAA         240                                                                           - ATTCACAGAC TCTCAAGATT TTAAAAAACT GTTTAACAAG GTCCCTATTG TT - #ACAGATGG         300                                                                           - AAGGGTCAAA CTTAATAAAG GATATTTGTT CGACTTTGTG ATTAGTTTGA TG - #CGATTCAA         360                                                                           - AAAAGAATCC TCTCTAGCTA CCACCGCAAT AGATCCTATT AGATACATAG AT - #CCTCGTCG         420                                                                           - CGATATCGCA TTTTCTAACG TGATGGATAT ATTAAAGTCG AATAAAGTGA AC - #AATAATTA         480                                                                           - ATTCTTTATT GTCATCATGT AATTAACTAG CTACCCGGGA GATCTCTCGA GC - #TGCAGAAG         540                                                                           - CTTATAAAAA TCACAAGTCT CTGTCACTTT TTTTGTCTAG TTTTTTTTTC TC - #CTCTTGGT         600                                                                           - TCAGACGTTC TCTTCTTCGT CGGAGTCTTT CAAGTGTCGG TAGCCGTTTT TG - #CGGTGTCG         660                                                                           - CAGTCGGTCT AGCAGGTTGG GCTTCTGTCC CTTGTCCTGC GTGCCAGTCT GT - #CCGTCCAA         720                                                                           - AGAATCTGTA CCGTTCTCGT GCGCTCGCTG CTCTGCGTCC AGACGGACCA GG - #GCCAGAAG         780                                                                           - CATCTGGTAA GCCTGCTCGT TGGTGTAAGG CGGAGCCGCC GTGGATGCAT CA - #GACGACGG         840                                                                           - TGGTCCCGGT CCTTTGCGAC CAGAATTATA AACACTTTCC TCGTAGGAAG GC - #GGAGCCTG         900                                                                           - TAACGACGTG TCTTTGGTGT TGCCCGACGT CACGGTGGTC CCGTCGGCGG AC - #ACCAGATA         960                                                                           - GGGAAAGAGG TTCTGCAGCG GCTGCATGCA GAGACGCCGC TGTCGAGTAT AG - #ATCAAATA        1020                                                                           - AATGATAATG ACGACGGCTA TGGCCACGAG GATGATGGTG AAGGCTCCGA AG - #GGGTTTTT        1080                                                                           - GAGGAAGGTG GCAACGCCTT CGACCACGGA GGCCACCGCG CCACCCACGG CC - #CCAATGGC        1140                                                                           - TACGCCAACG GCCTTTCCCG CGGCGCCCAG GCCGCTCATG AGGTCGTCCA GA - #CCCTTGAG        1200                                                                           - GTAGGGCGGC AGCGGGTCGA CTACCTTGTC CTCCACGTAC TTTACCCGCT GC - #TTATACGA        1260                                                                           - ATTGAACTCG CGCATGATCT CCTCGAGATC AAAAACGTTG CTGGAACGCA AT - #TCTTTCTG        1320                                                                           - CGAGTAAAGT TCCAGTACCC TGAAGTCGGT GTTTTCCAGC GGGTCGATGT CT - #AGGGCGAT        1380                                                                           - CATGCTGTCG ACGGTGGAGA TGCTGCTGAG GTCAATCATG CGTTTGAAGA GG - #TAGTCCAC        1440                                                                           - GTACTCGTAG GCCGAGTTGC CGGCGATGAA GATCTTGAGG CTGGGAAGCT GA - #CATTCCTC        1500                                                                           - AGTGCGGTGG TTGCCCAACA GGATTTCGTT ATCCTCGCCC AGTTGACCGT AC - #TGCACGTA        1560                                                                           - CGAGCTGTTG GCGAAATTAA AGATGACCAC TGGTCGTGAG TAGCAGCGTC CT - #GGCGATTC        1620                                                                           - CTTCACATTC ATATCACGCA GCACCTTGAC GCTGGTTTGG TTAATGGTCA CG - #CAGCTGGC        1680                                                                           - CAGACCCAGG ACATCACCCA TGAAACGCGC GGCAATCGGT TTGTTGTAGA TG - #GCCGAGAG        1740                                                                           - AATAGCTGAC GGGTTGATCT TGCTAAGTTC CTTGAAGACC TCTAGGGTGC GC - #CGTTGATC        1800                                                                           - CACACACCAG GCTTCTGCGA TTTCGGCCAG CGCCCGGTTG ATGTAACCGC GC - #AACGTGTC        1860                                                                           - ATAGGTGAAC TGCAGCTGGG CGTAGACCAG ATTGTGCACC GACTCCATGT TG - #GATAAATG        1920                                                                           - AGTTGCATTG TTGCCATCTG TACTTCTTTT GGTTCTATTA TGAGTAAGAT TC - #AGACTGGA        1980                                                                           - GCGGTTGGCC AAACGTTCGA GTTCCACCAG AGATTTTTGC TTGATACCTT GC - #CAGAACAC        2040                                                                           - CACCAAACCA CCAGTGGTTT CAAAGACGGA CACGTTTCCA TATTTTTCAT AT - #GTTTGATT        2100                                                                           - GTATGAAGTA TTGAAAATCT GCTGTAACTT ATTTATGGCC TCATCACGTA CA - #CAGTCCAG        2160                                                                           - CGCAGAGTCG GACATGTTCA CCTCTTGCTT CTTAGATAAG AAAGTGGCGG TC - #ATTTTGGC        2220                                                                           - AGAAGAAAAG TGATACGAGT CCTCGGCTTC GGAACGAATG GTGCGTTCCG AG - #GCTTCCCA        2280                                                                           - GAAAGTGAGT TGACAAGTAA CATTCTTCTC GTCCTGTATA TCCCAGGAGA TC - #ACTGAGTC        2340                                                                           - CGCACGTTCA AGAAAAGCCA CCAACCTGTG GGTCTCTAAC GCAGAATTCG GT - #CTTTCAAA        2400                                                                           - GTCGGAGACG ATAGTGTAGT TCGGAAAAAT GAAAAACTTG TCGGCGTTTT CT - #CCAAAATA        2460                                                                           - GCTGGCATTG CGATTAGTTC CGTTGTAGAA AGGAGAAATG TCAACCACAT CA - #CCCGTGGA        2520                                                                           - AGTTGCGAAA AAATGATAGG GATACTTGGA GCGCGCAGTA GTGATGGTCA CC - #ATACAATT        2580                                                                           - CAGATTACAG GTCTCACGAT AGAGCCAGGT GCTGCCGCGG CTGTGCCATT GA - #TCCTTGAC        2640                                                                           - CGTCACGTAA CGGGTACTGT GGGTGTTGGA ATAATCGTCG GGCATTAATT GC - #ATGGTTTT        2700                                                                           - GTTTTCATAG CTGTCCCTAT GATAAGCCAC GAAAACCGTG CCTGCTATAA CG - #CGGCTGTA        2760                                                                           - GGAACTGTAG CACTGACTGT GACTGTTGAT ATGATGAATC TCCCACATAG GA - #GGCGCCAC        2820                                                                           - GTATTCCGTG TTGCTGCCCA GCAGATAAGT GGTGTGGATG TAAGCGTAGC TA - #CGACGAAA        2880                                                                           - CGTCAAAACC TTCTGGTAGA CTCGTACCTT AAAGGTGTGC GCGACGATGT TG - #CGTTTGTA        2940                                                                           - GACCACCATG ATGCCCTCGT CCAGGTCTTC ATTGATGGGC TTCATCGAGG TG - #CAGACGAT        3000                                                                           - ATTACGTTCA AAGCGAATAA GATCCGTACC CTGTGCCATA GAACACACGC GA - #TAGGGGTA        3060                                                                           - CTTGGTGGTG TTGACCCCCA CCACATCTCC GTACTTGAGG GTAGTGTTGT AG - #ATGGTCTC        3120                                                                           - GTTAACACCA TGGCTGACCG TTTGGGAAGA AGTTACGCGT TGAGAGACTG AA - #CCGGATCG        3180                                                                           - AGAATGAGCA GCAGACGTCG TATGAGAGGA ATGGTGACTG TGAGTAGCAG AA - #GTTCCACG        3240                                                                           - AGTAGAAGAT GAGGAAACCG CAGCACCCAG ACAGACGATA CACAAGTTAA CG - #CAGACTAC        3300                                                                           - CAGGCACCAG ATCCTGGATT CCATTACGAT ACAAACTTAA CGGATATCGC GA - #TAATGAAA        3360                                                                           - TAATTTATGA TTATTTCTCG CTTTCAATTT AACACAACCC TCAAGAACCT TT - #GTATTTAT        3420                                                                           - TTTCACTTTT AAGTATAGAA TAAAGAAGCT TGCATGCCAC GCGTCTCGAG GG - #CCCCTGCA        3480                                                                           - GGTCGACTCT AGAGGATCCT TCTTTATTCT ATACTTAAAA AGTGAAAATA AA - #TACAAAGG        3540                                                                           - TTCTTGAGGG TTGTGTTAAA TTGAAAGCGA GAAATAATCA TAAATTATTT CA - #TTATCGCG        3600                                                                           - ATATCCGTTA AGTTTGTATC GTAATGTGCC GCCGCCCGGA TTGCGGCTTC TC - #TTTCTCAC        3660                                                                           - CTGGACCGGT GGCACTGCTG TGGTGTTGCC TTCTGCTGCC CATCGTTTCC TC - #AGCCACCG        3720                                                                           - TCAGCGTCGC TCCTACCGTC GCCGAGAAAG TTCCCGCGGA GTGCCCCGAA CT - #AACGCGTC        3780                                                                           - GATGCCTGTT GGGTGAGGTG TTTCAGGGTG ACAAGTATGA AAGTTGGCTG CG - #CCCGTTGG        3840                                                                           - TGAATGTTAC CAGACGCGAT GGCCCGCTAT CGCAACTTAT TCGTTACCGT CC - #CGTTACGC        3900                                                                           - CGGAGGCCGC CAACTCCGTG CTGTTGGACG ATGCTTTCCT GGACACTCTG GC - #CCTGCTGT        3960                                                                           - ACAACAATCC GGATCAATTG CGGGCCTTGC TGACGCTGTT GAGCTCGGAC AC - #AGCGCCGC        4020                                                                           - GCTGGATGAC GGTGATGCGC GGTTACAGCG AGTGCGGCGA TGGCTCGCCG GC - #CGTGTACA        4080                                                                           - CGTGCGTGGA CGACCTGTGC CGCGGCTACG ACCTCACGCG ACTGTCATAC GG - #GCGCAGCA        4140                                                                           - TCTTCACGGA ACACGTGTTA GGCTTCGAGC TGGTGCCACC GTCTCTCTTT AA - #CGTGGTGG        4200                                                                           - TGGCCATACG CAACGAAGCC ACGCGTACCA ACCGCGCCGT GCGTCTGCCC GT - #GAGCACCG        4260                                                                           - CTGCCGCGCC CGAGGGCATC ACGCTCTTTT ACGGCCTGTA CAACGCAGTG AA - #GGAATTCT        4320                                                                           - GCCTGCGTCA CCAGCTGGAC CCGCCGCTGC TACGCCACCT AGATAAATAC TA - #CGCCGGAC        4380                                                                           - TGCCGCCCGA GCTGAAGCAG ACGCGCGTCA ACCTGCCGGC TCACTCGCGC TA - #TGGCCCTC        4440                                                                           - AAGCAGTGGA TGCTCGCTAA TTTTTATAGA TCCTGATCCT TTTTCTGGGT AA - #GTAATACG        4500                                                                           - TCAAGGAGAA AACGAAACGA TCTGTAGTTA GCGGCCGCCT AATTAACTAA TA - #TTATATTT        4560                                                                           - TTTATCTAAA AAACTAAAAA TAAACATTGA TTAAATTTTA ATATAATACT TA - #AAAATGGA        4620                                                                           - TGTTGTGTCG TTAGATAAAC CGTTTATGTA TTTTGAGGAA ATTGATAATG AG - #TTAGATTA        4680                                                                           - CGAACCAGAA AGTGCAAATG AGGTCGCAAA AAAACTGCCG TATCAAGGAC AG - #TTAAAACT        4740                                                                           - ATTACTAGGA GAATTATTTT TTCTTAGTAA GTTACAGCGA CACGGTATAT TA - #GATGGTGC        4800                                                                           - CACCGTAGTG TATATAGGAT CGGCTCCTGG TACACATATA CGTTATTTGA GA - #GATCATTT        4860                                                                           - CTATAATTTA GGAATGATTA TCAAATGGAT GCTAATTGAC GGACGCCATC AT - #GATCCTAT        4920                                                                           - TTTAAATGGA TTGCGTGATG TGACTCTAGT GACTCGGTTC GTTGATGAGG AA - #TATCTACG        4980                                                                           - ATCCATCAAA AAACAACTGC ATCCTTCTAA GATTATTTTA ATTTCTGATG TG - #AGATCCAA        5040                                                                           - ACGAGGAGGA AATGAACCTA GTACGGCGGA TTTACTAAGT AATTACGCTC TA - #CAAAATGT        5100                                                                           - CATGATTAGT ATTTTAAACC CCGTGGCGTC TAGTCTTAAA TGGAGATGCC CG - #TTTCCAGA        5160                                                                           - TCAATGGATC AAGGACTTTT ATATCCCACA CGGTAATAAA ATGTTACAAC CT - #TTTGCTCC        5220                                                                           #   5234                                                                       - (2) INFORMATION FOR SEQ ID NO:74:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 56 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74:                                 - GCCTCATCGC TGCTGGATAT CCGTTAAGTT TGTATCGTAA TGGAATCCAG GA - #TCTG              56                                                                           - (2) INFORMATION FOR SEQ ID NO:75:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 40 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:75:                                 #    40            TTTT ATAAGCTTCG TAAGCTGTCA                                  - (2) INFORMATION FOR SEQ ID NO:76:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 55 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:76:                                 - AGCTTCTTTA TTCTATACTT AAAAAGTGAA AATAAATACA AAGGTTCTTG AG - #GGT               55                                                                           - (2) INFORMATION FOR SEQ ID NO:77:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 73 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:77:                                 - TGTGTTAAAT TGAAAGCGAG AAATAATCAT AAATTATTTC ATTATCGCGA TA - #TCCGTTAA          60                                                                           #      73                                                                      - (2) INFORMATION FOR SEQ ID NO:78:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 56 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:78:                                 - TTATTAGTAT TTAATAAAGT AATAGCGCTA TAGGCAATTC AAACATAGCA TG - #AGCT              56                                                                           - (2) INFORMATION FOR SEQ ID NO:79:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 72 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:79:                                 - AGAAATAAGA TATGAATTTT TCACTTTTAT TTATGTTTCC AAGAACTCCC AA - #CACAATTT          60                                                                           #       72                                                                     - (2) INFORMATION FOR SEQ ID NO:80:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 14 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:80:                                 #     14                                                                       - (2) INFORMATION FOR SEQ ID NO:81:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 22 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:81:                                 #                 22CTG CA                                                     - (2) INFORMATION FOR SEQ ID NO:82:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 29 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82:                                 #            29    TATT TTATTTCAG                                              - (2) INFORMATION FOR SEQ ID NO:83:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 61 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:83:                                 - TATCTGAATT CCTGCAGCCC GGGTTTTTAT AGCTAATTAG TCAAATGTGA GT - #TAATATTA          60                                                                           #               61                                                             - (2) INFORMATION FOR SEQ ID NO:84:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 66 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84:                                 - TCGCTGAATT CGATATCAAG CTTATCGATT TTTATGACTA GTTAATCAAA TA - #AAAAGCAT          60                                                                           #           66                                                                 - (2) INFORMATION FOR SEQ ID NO:85:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 30 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:85:                                 #           30     CATC AGTATCTAAC                                             - (2) INFORMATION FOR SEQ ID NO:86:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 37 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:86:                                 #      37          CAGA TATTTGTTAG CTTCTGC                                     - (2) INFORMATION FOR SEQ ID NO:87:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 33 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:87:                                 #         33       CCTA CCTACTACCT ACG                                         - (2) INFORMATION FOR SEQ ID NO:88:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 29 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88:                                 #            29    GACA ATAACATAG                                              - (2) INFORMATION FOR SEQ ID NO:89:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 30 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89:                                 #           30     TACT GGGTTACAAC                                             - (2) INFORMATION FOR SEQ ID NO:90:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 17 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:90:                                 #   17             C                                                           - (2) INFORMATION FOR SEQ ID NO:91:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 15 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:91:                                 #    15                                                                        - (2) INFORMATION FOR SEQ ID NO:92:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 39 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:92:                                 #    39            TTAA TTAGTTATTA GACAAGGTG                                   - (2) INFORMATION FOR SEQ ID NO:93:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 41 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:93:                                 #   41             ACGA TACAAACTTA AGCGGATATC G                                - (2) INFORMATION FOR SEQ ID NO:94:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 45 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94:                                 #45                GCCG CTCATTAGAC AAGCGAATGA GGGAC                            - (2) INFORMATION FOR SEQ ID NO:95:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 62 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:95:                                 - AGATCTCCCG GGCTCGAGTA ATTAATTAAT TTTTATTACA CCAGAAAAGA CG - #GCTTGAGA          60                                                                           #              62                                                              - (2) INFORMATION FOR SEQ ID NO:96:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 64 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96:                                 - TAATTACTCG AGCCCGGGAG ATCTAATTTA ATTTAATTTA TATAACTCAT TT - #TTTGAATA          60                                                                           #             64                                                               - (2) INFORMATION FOR SEQ ID NO:97:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 46 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:97:                                 #                 46GCT TTAAATGGAC GGAACTCTTT TCCCCC                           - (2) INFORMATION FOR SEQ ID NO:98:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 62 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:98:                                 - GATCTTTTGT TAACAAAAAC TAATCAGCTA TCGCGAATCG ATTCCCGGGG GA - #TCCGGTAC          60                                                                           #              62                                                              - (2) INFORMATION FOR SEQ ID NO:99:                                            -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 62 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:99:                                 - TCGAGGGTAC CGGATCCCCC GGGAATCGAT TCGCGATAGC TGATTAGTTT TT - #GTTAACAA          60                                                                           #              62                                                              - (2) INFORMATION FOR SEQ ID NO:100:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 46 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:100:                                #                 46GCG GCGTCTCTGC ATGCAGCCGC TGCAGA                           - (2) INFORMATION FOR SEQ ID NO:101:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 46 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:101:                                #                 46CAT GCAGAGACGC CGCTGTCGAG TCCATG                           - (2) INFORMATION FOR SEQ ID NO:102:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 33 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:102:                                #         33       CACC TATGACACGT TGC                                         - (2) INFORMATION FOR SEQ ID NO:103:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 37 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:103:                                #      37          TCCA GACCCTTGAG GTAGGGC                                     - (2) INFORMATION FOR SEQ ID NO:104:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 48 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:104:                                #                48CTGG ACGACACTCG ACAGCGGCGT CTCTGCAT                         - (2) INFORMATION FOR SEQ ID NO:105:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 12 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:105:                                #       12                                                                     - (2) INFORMATION FOR SEQ ID NO:106:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 12 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:106:                                #       12                                                                     - (2) INFORMATION FOR SEQ ID NO:107:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:107:                                # 20               GCGT                                                        - (2) INFORMATION FOR SEQ ID NO:108:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 25 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:108:                                #               25 GTTG TAGAT                                                  - (2) INFORMATION FOR SEQ ID NO:109:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:109:                                # 20               CGGC                                                        - (2) INFORMATION FOR SEQ ID NO:110:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 39 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:110:                                #    39            TAAG ATCTACAGAT GGCAACAAT                                   - (2) INFORMATION FOR SEQ ID NO:111:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 33 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:111:                                #         33       CAGG CCTCCCCTCC TAC                                         - (2) INFORMATION FOR SEQ ID NO:112:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 29 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:112:                                #            29    GTCT TGAGCATGC                                              - (2) INFORMATION FOR SEQ ID NO:113:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 29 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:113:                                #            29    TTTT ATCTCGAGA                                              - (2) INFORMATION FOR SEQ ID NO:114:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 37 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:114:                                #      37          AATC AGCATGTCTT GAGCATG                                     - (2) INFORMATION FOR SEQ ID NO:115:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 46 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:115:                                #                 46GGA AGAATATGAT AATATTTTGG GATTTC                           - (2) INFORMATION FOR SEQ ID NO:116:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 42 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:116:                                #  42              AATT ACAATATAAA ATGCGGCCCG GG                               - (2) INFORMATION FOR SEQ ID NO:117:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 34 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:117:                                #        34        TTTA TATTGTAATT ATAT                                        - (2) INFORMATION FOR SEQ ID NO:118:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 54 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:118:                                - ATTTTCAATT TTGAAATCCC AAAATATTAT CATATTCTTC CCAATAAACT CG - #AG                54                                                                           - (2) INFORMATION FOR SEQ ID NO:119:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 35 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:119:                                #       35         TCCC CTCCTACCTC ATCGT                                       - (2) INFORMATION FOR SEQ ID NO:120:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 34 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:120:                                #        34        TAAG TCTCTGTTGG TATC                                        - (2) INFORMATION FOR SEQ ID NO:121:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 66 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:121:                                - AGAAAAATCA GTTAGCTAAG ATCTCCCGGG CTCGAGGGTA CCGGATCCTG AT - #TAGTTAAT          60                                                                           #           66                                                                 - (2) INFORMATION FOR SEQ ID NO:122:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 70 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:122:                                - GATCACAAAA ATTAACTAAT CAGGATCCGG TACCCTCGAG CCCGGGAGAT CT - #TAGCTAAC          60                                                                           #        70                                                                    - (2) INFORMATION FOR SEQ ID NO:123:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 35 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:123:                                #       35         TGTT AAATGTTATA CTTTG                                       - (2) INFORMATION FOR SEQ ID NO:124:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 28 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:124:                                #             28   GTAC CACTTCAG                                               - (2) INFORMATION FOR SEQ ID NO:125:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 44 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:125:                                # 44               CTTA TAAAGATCTA AAATGCATAA TTTC                             - (2) INFORMATION FOR SEQ ID NO:126:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 35 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:126:                                #       35         TCTA AACTAGGAAT AGATG                                       - (2) INFORMATION FOR SEQ ID NO:127:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 82 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:127:                                - GTACGTGACT AATTAGCTAT AAAAAGGATC CGGTACCCTC GAGTCTAGAA TC - #GATCCCGG          60                                                                           #                 82ATC AC                                                     - (2) INFORMATION FOR SEQ ID NO:128:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 82 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:128:                                - GGCCGTGATT AACTAGTCAT AAAAACCCGG GATCGATTCT AGACTCGAGG GT - #ACCGGATC          60                                                                           #                 82GTC AC                                                     - (2) INFORMATION FOR SEQ ID NO:129:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 70 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:129:                                - GATCTTAATT AATTAGTCAT CAGGCAGGGC GAGAACGAGA CTATCTGCTC GT - #TAATTAAT          60                                                                           #        70                                                                    - (2) INFORMATION FOR SEQ ID NO:130:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 70 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:130:                                - GATCCGTCGA CCTAATTAAT TAACGAGCAG ATAGTCTCGT TCTCGCCCTG CC - #TGATGACT          60                                                                           #        70                                                                    - (2) INFORMATION FOR SEQ ID NO:131:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 12 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:131:                                #       12                                                                     - (2) INFORMATION FOR SEQ ID NO:132:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 78 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:132:                                - ATAAAAATTA GCTACTCAGG TACCCTGCAG TCGCGAGGAT CCGAATTCCC CG - #GGCTCGAG          60                                                                           #  78              AT                                                          - (2) INFORMATION FOR SEQ ID NO:133:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 78 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:133:                                - ATAAAAACTA ATTAATCACT CGAGCCCGGG GAATTCGGAT CCTCGCGACT GC - #AGGGTACC          60                                                                           #  78              AT                                                          - (2) INFORMATION FOR SEQ ID NO:134:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 35 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:134:                                #       35         TACT GGTCAGCCTT GCTTC                                       - (2) INFORMATION FOR SEQ ID NO:135:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 42 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:135:                                #  42              TCGT AATGGAGTCC TCTGCCAAGA GA                               - (2) INFORMATION FOR SEQ ID NO:136:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 44 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:136:                                # 44               ATCC GTTAAGTTTG TATCGTAATG GAGT                             - (2) INFORMATION FOR SEQ ID NO:137:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 30 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:137:                                #           30     TCCT TCCTCAACAT                                             - (2) INFORMATION FOR SEQ ID NO:138:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 29 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:138:                                #            29    AGTG TAATGAAGC                                              - (2) INFORMATION FOR SEQ ID NO:139:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 46 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:139:                                #                 46AAA AATTACTGGT CAGCCTTGCT TCTAGT                           - (2) INFORMATION FOR SEQ ID NO:140:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 43 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:140:                                # 43               TGTA TCGTAATCTG CAGCCCGGGG GGG                              - (2) INFORMATION FOR SEQ ID NO:141:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 44 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:141:                                # 44               GATT ACGATACAAA CTTAACGGAT ATCG                             - (2) INFORMATION FOR SEQ ID NO:142:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 60 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:142:                                - CGCGAATTCT CGCGATATCC GTTAAGTTTG TATCGTAATG AAACAGATTA AG - #GTTCGAGT          60                                                                           - (2) INFORMATION FOR SEQ ID NO:143:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 27 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:143:                                #             27   ATGG CCTGACT                                                - (2) INFORMATION FOR SEQ ID NO:144:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 39 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:144:                                #    39            TTAA TTAGTCATCA GGCAGGGCG                                   - (2) INFORMATION FOR SEQ ID NO:145:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 40 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:145:                                #    40            ACGA TACAAACTTA ACGGATATCG                                  - (2) INFORMATION FOR SEQ ID NO:146:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 27 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:146:                                #             27   TTCT ATACTTA                                                - (2) INFORMATION FOR SEQ ID NO:147:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 72 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:147:                                - AATTCTCGCG ATATCCGTTA AGTTTGTATC GTAATGACGA CGTTCCTGCA GA - #CTATGTTG          60                                                                           #       72                                                                     - (2) INFORMATION FOR SEQ ID NO:148:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 68 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:148:                                - AACCTCCTTC CTCAACATAG TCTGCAGGAA CGTCGTCATT ACGATACAAA CT - #TAACGGAT          60                                                                           #          68                                                                  - (2) INFORMATION FOR SEQ ID NO:149:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 39 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:149:                                #    39            ACGA TTGTTCATGA TGGCAAGAT                                   - (2) INFORMATION FOR SEQ ID NO:150:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 68 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:150:                                - CCCGGGGGAT CCCTCGAGGG TACCAAGCTT AATTAATTAA ATATTAGTAT AA - #AAAGTGAT          60                                                                           #          68                                                                  - (2) INFORMATION FOR SEQ ID NO:151:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 77 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:151:                                - AAGCTTGGTA CCCTCGAGGG ATCCCCCGGG TAGCTAGCTA ATTTTTCTTT TA - #CGTATTAT          60                                                                           #   77             C                                                           - (2) INFORMATION FOR SEQ ID NO:152:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 39 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:152:                                #    39            GTAT TTTTAAAACT TCTAACACC                                   - (2) INFORMATION FOR SEQ ID NO:153:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 62 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:153:                                - GATTATCGCG ATATCCGTTA AGTTTGTATC GTAATGGCAT CCGTACTGGG TC - #CCATTTCG          60                                                                           #              62                                                              - (2) INFORMATION FOR SEQ ID NO:154:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 47 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:154:                                #                47CATA AAAATCAACC TCGGTGCTTT TTGGGCG                          - (2) INFORMATION FOR SEQ ID NO:155:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 29 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:155:                                #            29    TCAG TCGCCTACA                                              - (2) INFORMATION FOR SEQ ID NO:156:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 29 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:156:                                #            29    AAAA AGAAGAGCG                                              - (2) INFORMATION FOR SEQ ID NO:157:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 61 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:157:                                - GATTATCGCG ATATCCGTTA AGTTTGTATC GTAATGGAGT CGCGCGGTCG CC - #GTTGTCCC          60                                                                           #               61                                                             - (2) INFORMATION FOR SEQ ID NO:158:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 17 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:158:                                #   17             C                                                           - (2) INFORMATION FOR SEQ ID NO:159:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 42 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:159:                                #  42              CCGC CTATCAAAAG TCTTAATGAG TT                               - (2) INFORMATION FOR SEQ ID NO:160:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 72 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:160:                                - GAATTCCTCG AGCTGCAGCC CGGGTTTTTA TAGCTAATTA GTCATTTTTT CG - #TAAGTAAG          60                                                                           #       72                                                                     - (2) INFORMATION FOR SEQ ID NO:161:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 72 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:161:                                - CCCGGGCTGC AGCTCGAGGA ATTCTTTTTA TTGATTAACT AGTCAAATGA GT - #ATATATAA          60                                                                           #       72                                                                     - (2) INFORMATION FOR SEQ ID NO:162:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 45 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:162:                                #45                TAAA TACAAGTTTG ATTAAACTTA AGTTG                            - (2) INFORMATION FOR SEQ ID NO:163:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 26 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:163:                                #              26  GAGA AAAGCC                                                 - (2) INFORMATION FOR SEQ ID NO:164:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 32 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:164:                                #          32      GAAG GGATTGTAAG CC                                          - (2) INFORMATION FOR SEQ ID NO:165:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 28 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:165:                                #             28   AGTC TCGTCTCC                                               - (2) INFORMATION FOR SEQ ID NO:166:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 30 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:166:                                #           30     TGCC CGCGGACAAC                                             - (2) INFORMATION FOR SEQ ID NO:167:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 99 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:167:                                - TTCGAATTCG CTAGCTTTAT TGGGAAGAAT ATGATAATAT TTTGGGATTT CA - #AAATTGAA          60                                                                           #    99            ATAA AATGAGTTTG CAGTTTATC                                   - (2) INFORMATION FOR SEQ ID NO:168:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 28 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:168:                                #             28   GTTG AACAGCAC                                               - (2) INFORMATION FOR SEQ ID NO:169:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 44 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:169:                                # 44               ATAA AAACTATTCC TCCGTGTTCT TAAT                             - (2) INFORMATION FOR SEQ ID NO:170:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 28 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:170:                                #             28   AGCT AAGTTATC                                               - (2) INFORMATION FOR SEQ ID NO:171:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 30 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:171:                                #           30     GCCG CCCGGATTGC                                             - (2) INFORMATION FOR SEQ ID NO:172:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 33 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:172:                                #         33       ATCC ACTGCTTGAG GGC                                         - (2) INFORMATION FOR SEQ ID NO:173:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 54 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:173:                                - TCCAAGCTTA GATCTATAAA AATTAGCGAG CATCCACTGC TTGAGGGCCA TA - #GC                54                                                                           - (2) INFORMATION FOR SEQ ID NO:174:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 32 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:174:                                #          32      GCTG TTGAGCTCGG AC                                          - (2) INFORMATION FOR SEQ ID NO:175:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 58 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:175:                                - CGCGAATTCT CGCGATATCC GTTAAGTTTG TATCGTAATG TGCCGCCGCC CG - #GATTGC            58                                                                           - (2) INFORMATION FOR SEQ ID NO:176:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 34 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:176:                                #        34        GCGG CGCTGTGTCC GAGC                                        - (2) INFORMATION FOR SEQ ID NO:177:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 6749 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:177:                                - GAGCTCGCGG CCGCCTATCA AAAGTCTTAA TGAGTTAGGT GTAGATAGTA TA - #GATATTAC          60                                                                           - TACAAAGGTA TTCATATTTC CTATCAATTC TAAAGTAGAT GATATTAATA AC - #TCAAAGAT         120                                                                           - GATGATAGTA GATAATAGAT ACGCTCATAT AATGACTGCA AATTTGGACG GT - #TCACATTT         180                                                                           - TAATCATCAC GCGTTCATAA GTTTCAACTG CATAGATCAA AATCTCACTA AA - #AAGATAGC         240                                                                           - CGATGTATTT GAGAGAGATT GGACATCTAA CTACGCTAAA GAAATTACAG TT - #ATAAATAA         300                                                                           - TACATAATGG ATTTTGTTAT CATCAGTTAT ATTTAACATA AGTACAATAA AA - #AGTATTAA         360                                                                           - ATAAAAATAC TTACTTACGA AAAAATGACT AATTAGCTAT AAAAACCCAA CA - #AAAACTAA         420                                                                           - TCAGCTATCG GGGTTAATTA ATTAGTTATT AGACAAGGTG AAAACGAAAC TA - #TTTGTAGC         480                                                                           - TTAATTAATT AGAGCTTCTT TATTCTATAC TTAAAAAGTG AAAATAAATA CA - #AAGGTTCT         540                                                                           - TGAGGGTTGT GTTAAATTGA AAGCGAGAAA TAATCATAAA TTATTTCATT AT - #GGCGATAT         600                                                                           - CCGTTAAGTT TGTATCGTAA TGGAGTCGCG CGGTCGCCGT TGTCCCGAAA TG - #ATATCCGT         660                                                                           - ACTGGGTCCC ATTTCGGGGC ACGTGCTGAA AGCCGTGTTT AGTCGCGGCG AC - #ACGCCGGT         720                                                                           - GCTGCCGCAC GAGACGCGAC TCCTGCAGAC GGGTATCCAC GTGCGCGTGA GC - #CAGCCCTC         780                                                                           - GCTGATCCTG GTGTCGCAGT ACACGCCCGA CTCGACGCCA TGCCACCGCG GC - #GACAATCA         840                                                                           - GCTGCAGGTG CAGCACACGT ACTTTACGGG CAGCGAGGTG GAGAACGTGT CG - #GTCAACGT         900                                                                           - GCACAACCCC ACGGGCCGGA GCATCTGCCC CAGCCAAGAG CCCATGTCGA TC - #TATGTGTA         960                                                                           - CGCGCTGCCG CTCAAGATGC TGAACATCCC CAGCATCAAC GTGCACCACT AC - #CCGTCGGC        1020                                                                           - GGCCGAGCGC AAACACCGAC ACCTGCCCGT AGCTGACGCT GTGATTCACG CG - #TCGGGCAA        1080                                                                           - GCAGATGTGG CAGGCGCGTC TCACGGTCTC GGGACTGGCC TGGACGCGTC AG - #CAGAACCA        1140                                                                           - GTGGAAAGAG CCCGACGTCT ACTACACGTC AGCGTTCGTG TTTCCCACCA AG - #GACGTGGC        1200                                                                           - ACTGCGGCAC GTGGTGTGCG CGCACGAGCT GGTTTGCTCC ATGGAGAACA CG - #CGCGCAAC        1260                                                                           - CAAGATGCAG GTGATAGGTG ACCAGTACGT CAAGGTGTAC CTGGAGTCCT TC - #TGCGAGGA        1320                                                                           - CGTGCCCTCC GGCAAGCTCT TTATGCACGT CACGCTGGGC TCTGACGTGG AA - #GAGGACCT        1380                                                                           - GACGATGACC CGCAACCCGC AACCCTTCAT GCGCCCCCAC GAGCGCAACG GC - #TTTACGGT        1440                                                                           - GTTGTGTCCC AAAAATATGA TAATCAAACC GGGCAAGATC TCGCACATCA TG - #CTGGATGT        1500                                                                           - GGCTTTTACC TCACACGAGC ATTTTGGGCT GCTGTGTCCC AAGAGCATCC CG - #GGCCTGAG        1560                                                                           - CATCTCAGGT AACCTATTGA TGAACGGGCA GCAGATCTTC CTGGAGGTGC AA - #GCGATACG        1620                                                                           - CGAGACCGTG GAACTGCGTC AGTACGATCC CGTGGCTGCG CTCTTCTTTT TC - #GATATCGA        1680                                                                           - CTTGCTGCTG CAGCGCGGGC CTCAGTACAG CGAACACCCC ACCTTCACCA GC - #CAGTATCG        1740                                                                           - CATCCAGGGC AAGCTTGAGT ACCGACACAC CTGGGACCGG CACGACGAGG GT - #GCCGCCCA        1800                                                                           - GGGCGACGAC GACGTCTGGA CCAGCGGATC GGACTCCGAC GAGGAACTCG TA - #ACCACCGA        1860                                                                           - GCGCAAGACG CCCCGCGTTA CCGGCGGCGG CGCCATGGCG GGCGCCTCCA CT - #TCCGCGGG        1920                                                                           - CCGCAAACGC AAATCAGCAT CCTCGGCGAC GGCGTGCACG GCGGGCGTTA TG - #ACACGCGG        1980                                                                           - CCGCCTTAAG GCCGAGTCCA CCGTCGCGCC CGAAGAGGAC ACCGACGAGG AT - #TCCGACAA        2040                                                                           - CGAAATCCAC AATCCGGCCG TGTTCACCTG GCCGCCCTGG CAGGCCGGCA TC - #CTGGCCCG        2100                                                                           - CAACCTGGTG CCCATGGTGG CTACGGTTCA GGGTCAGAAT CTGAAGTACC AG - #GAGTTCTT        2160                                                                           - CTGGGACGCC AACGACATCT ACCGCATCTT CGCCGAATTG GAAGGCGTAT GG - #CAGCCCGC        2220                                                                           - TGCGCAACCC AAACGTCGCC GCCACCGGCA AGACGCCTTG CCCGGGCCAT GC - #ATCGCCTC        2280                                                                           - GACGCCCAAA AAGCACCGAG GTTGATTTTT ATGGATCCGG TACCCTCGAG GA - #ATTCTAGC        2340                                                                           - TTTATTGGGA AGATATGATA ATATTTTGGG ATTTCAAAAT TGAAAATATA TA - #ATTACAAT        2400                                                                           - ATAAAATGAG TTTGCAGTTT ATCGGTCTAC AGCGGCGCGA TGTGGTGGCC CT - #GGTCAACT        2460                                                                           - TTCTGCGCCA TCTCACGCAA AAGCCCGACG TGGATCTCGA GGCACACCCC AA - #GATCCTGA        2520                                                                           - AAAAATGTGG CGAAAAACGC CTGCACCGGC GTACGGTGCT GTTCAACGAG CT - #CATGCTTT        2580                                                                           - GGTTGGGATA CTACCGCGAG CTGCGTTTCC ACAACCCCGA CCTCTCCTCG GT - #TCTCGAGG        2640                                                                           - AGTTCGAGGT GCGTTGCGCG GCCGTGGCGC GTCGCGGCTA CACTTACCCG TT - #CGGTGATC        2700                                                                           - GTGGTAAGGC GCGTGACCAC CTGGCTGTGC TAGACCGTAC CGAATTCGAT AC - #GGACGTAC        2760                                                                           - GCCACGATGC TGAGATTGTG GAGCGCGCGC TCGTAAGCGC GGTCATTCTG GC - #CAAGATGT        2820                                                                           - CGGTGCGCGA GACGCTGGTC ACAGCCATCG GCCAGACGGA ACCCATCGCT TT - #TGTGCACC        2880                                                                           - TCAAGGATAC GGAGGTGCAG CGCATTGAAG AAAACCTGGA GGGTGTGCGC CG - #TAACATGT        2940                                                                           - TCTGCGTGAA ACCGCTCGAC CTTAACCTGG ACCGGCACGC CAACACGGCG CT - #GGTCAACG        3000                                                                           - CCGTCAACAA GCTCGTGTAC ACGGGCCGTC TCATCATGAA CGTGCGCAGG TC - #TTGGGAGG        3060                                                                           - AGCTGGAGCG CAAATGTCTG GCGCGCATTC AGGAGCGCTG CAAGCTGCTG GT - #CAAGGAGC        3120                                                                           - TGCGCATGTG CCTTTCCTTT GATTCCAACT ACTGTCGCAA TATCCTCAAA CA - #CGCCGTGG        3180                                                                           - AAAACGGTGA CTCGGCCGAC ACGCTGCTGG AGCTGCTCAT CGAGGACTTT GA - #CATCTACG        3240                                                                           - TGGACAGCTT CCCGCAGTCG GCGCACACCT TTTTGGGCGC GCGCCCGCCG TC - #GTTGGAGT        3300                                                                           - TTGACGATGA CGCCAATCTC CTCTCGCTCG GCGGCGGTTC AGCCTTCTCG TC - #GGTACCCA        3360                                                                           - AGAAACATGT CCCCACGCAG CCGCTGGACG GCTGGAGCTG GATCGCCAGT CC - #CTGGAAGG        3420                                                                           - GACACAAACC GTTCCGCTTC GAGGCCCATG GTTCTCTGGC ACCGGCCGCC GA - #CGCCCACG        3480                                                                           - CCGCCCGTTC GGCGCGCGTC GGCTATTACG ACGAAGAGGA AAAGCGTCGC GA - #GCGGCAGA        3540                                                                           - AACGGGTGGA CGACGAGGTG GTGCAGCGTG AGAAACAGCA GCTGAAGGCT TG - #GGAGGAGA        3600                                                                           - GGCAGCAGAA CCTGCAGCAA CGTCAGCAGC AACCGCCGCC CCCGACACGT AA - #ACCGGGCG        3660                                                                           - CCTCCCGGAG GCTCTTTGGC TCCAGTGCCG ATGAGGACGA CGACGATGAT GA - #TGACGAGA        3720                                                                           - AAAACATCTT TACGCCCATC AAGAAACCGG GAACTAGCGG CAAGGGCGCC GC - #TAGTGGCA        3780                                                                           - ACGGTGTTTC CAGCATTTTC AGCGGCATGT TATCCTCGGG CAGTCAGAAA CC - #GACCAGCG        3840                                                                           - GTCCCTTGAA CATCCCGCAG CAACAACAGC GTCACGCGGC TTTCAGTCTC GT - #CTCCCCGC        3900                                                                           - AGGTAACCAA GGCCAGCCCG GGAAGGGTCC GTCGGGACAG CGCGTGGGAC GT - #GAGGCCGC        3960                                                                           - TCACGGAGAC AAGAGGGGAT CTTTTCTCGG GCGACGAGGA TTCCGACAGC TC - #GGATGGCT        4020                                                                           - ATCCCCCCAA CCGTCAAGAT CCGCGTTTCA CCGACACGCT GGTGGACATC AC - #GGATACCG        4080                                                                           - AGACGAGCGC CAAACCGCCC GTCACCACCG CGTACAAGTT CGAGCAACCG AC - #GTTGACGT        4140                                                                           - TCGGCGCCGG AGTTAACGTC CCTGCTGGCG CCGGCGCTGC CATCCTCACG CC - #GACGCCTG        4200                                                                           - TCAATCCTTC CACGGCCCCC GCTCCGGCCC CGACACCTAC CTTCGCGGGT AC - #CCAAACCC        4260                                                                           - CGGTCAACGG TAACTCGCCC TGGGCTCCGA CGGCGCCGTT GCCCGGGGAT AT - #GAACCCCG        4320                                                                           - CCAACTGGCC GCGCGAACGC GCGTGGGCCC TCAAGAATCC TCACCTGGCT TA - #CAATCCCT        4380                                                                           - TCAGGATGCC TACGACTTCC ACGACTTCTC AAAACAACGT GTCCACCACC CC - #TCGGAGGC        4440                                                                           - CGTCGACTCC ACGCGCCGCG GTGACACAAA CAGCGTCTCA GAACGCCGCT GA - #TGAGGTTT        4500                                                                           - GGGCTTTAAG GGACCAAACT GCAGAGTCAC CGGTCGAAGA CAGCGAGGAG GA - #AGACGACG        4560                                                                           - ACTCCTCGGA CACCGGCTCC GTCGTCAGCC TGGGACACAC AACACCGTCG TC - #CGATTACA        4620                                                                           - ACGACGTCAT TTCGCCTCCC AGTCAGACGC CCGAGCAGTC GACGCCGTCC AG - #AATACGTA        4680                                                                           - AAGCTAAGTT ATCGTCTCCA ATGACGACGA CATCCACGAG CCAGAAACCG GT - #GCTGGGCA        4740                                                                           - AGCGAGTCGC GACGCCGCAC GCGTCCGCCC GAGCGCAGAC GGTGACGTCG AC - #ACCGGTTC        4800                                                                           - AGGGAAGGGT AGAGAAACAG GTATCGGGCA CGCCGTCGAC GGTACCCGCC AC - #GCTGTTGC        4860                                                                           - AACCTCAACC GGCTTCGTCT AAAACAACGT CATCAAGGAA CGTGACTTCT GG - #CGCGAGAA        4920                                                                           - CCTCTTCCGC TTCGGCTCGA CAGCCGTCAG CCTCGGCGTC CGTTTTGTCG CC - #CACGGAGG        4980                                                                           - ATGATGTCGT GTCCCCCGTC ACGTCGCCGC TGTCCATGCT TTCGTCAGCC TC - #TCCGTCCC        5040                                                                           - CGGCCAAGAG TGCCCCTCCG TCTCCGGTGA AAGGTCGGGG CAGCCGCGTC GG - #TGTTCCTT        5100                                                                           - CTTTGAAACC TACTTTGGGC GGCAAGGCGG TGGTAGGTCG ACCGCCCTCG GT - #ACCCGTGA        5160                                                                           - GCGGTAGCGC GCCGGGTCGC CTGTCCGGCA CCAGCCGGGC CGCCTCGACC AC - #GCCGACGT        5220                                                                           - ATCCCGCGGT AACCACCGTT TACCCACCGT CGTCTACGGC CAAAAGCAGC GT - #ATCGAATG        5280                                                                           - CGCCGCCTGT GGCCTCCCCC TCCATCCTGA AACCGGGGGC GAGCGCGGCT TT - #GCAATCAC        5340                                                                           - GCCGCTCGAC GGGGACCGCC GCCGTAGGTT CCCCCGTCAA GAGCACGACG GG - #CATGAAAA        5400                                                                           - CGGTGGCTTT CGACCTATCG TCGCCCCAGA AGAGCGGTAC GGGGCCGCAA CC - #GGGTTCTG        5460                                                                           - CCGGCATGGG GGGCGCCAAA ACGCCGTCGG ACGCCGTGCA GAACATCCTC CA - #AAAGATCG        5520                                                                           - AGAAGATTAA GAACACGGAG GAATAGTTTT TATTGCTAGA ATTCTTTTTA TT - #GATTAACT        5580                                                                           - AGTCAAATGA GTATATATAA TTGAAAAAGT AAAATATAAA TCATATAATA AT - #GAAACGAA        5640                                                                           - ATATCAGTAA TAGACAGGAA CTGGCAGATT CTTCTTCTAA TGAAGTAAGT AC - #TGCTAAAT        5700                                                                           - CTCCAAAATT AGATAAAAAT GATACAGCAA ATACAGCTTC ATTCAACGAA TT - #ACCTTTTA        5760                                                                           - ATTTTTTCAG ACACACCTTA TTACAAACTA ACTAAGTCAG ATGATGAGAA AG - #TAAATATA        5820                                                                           - AATTTAACTT ATGGGTATAA TATAATAAAG ATTCATGATA TTAATAATTT AC - #TTAACGAT        5880                                                                           - GTTAATAGAC TTATTCCATC AACCCCTTCA AACCTTTCTG GATATTATAA AA - #TACCAGTT        5940                                                                           - AATGATATTA AAATAGATTG TTTAAGAGAT GTAAATAATT ATTTGGAGGT AA - #AGGATATA        6000                                                                           - AAATTAGTCT ATCTTTCACA TGGAAATGAA TTACCTAATA TTAATAATTA TG - #ATAGGAAT        6060                                                                           - TTTTTAGGAT TTACAGCTGT TATATGTATC AACAATACAG GCAGATCTAT GG - #TTATGGTA        6120                                                                           - AAACACTGTA ACGGGAAGCA GCATTCTATG GTAACTGGCC TATGTTTAAT AG - #CCAGATCA        6180                                                                           - TTTTACTCTA TAAACATTTT ACCACAAATA ATAGGATCCT CTAGATATTT AA - #TATTATAT        6240                                                                           - CTAACAACAA CAAAAAAATT TAACGATGTA TGGCCAGAAG TATTTTCTAC TA - #ATAAAGAT        6300                                                                           - AAAGATAGTC TATCTTATCT ACAAGATATG AAAGAAGATA ATCATTTAGT AG - #TAGCTACT        6360                                                                           - AATATGGAAA GAAATGTATA CAAAAACGTG GAAGCTTTTA TATTAAATAG CA - #TATTACTA        6420                                                                           - GAAGATTTAA AATCTAGACT TAGTATAACA AAACAGTTAA ATGCCAATAT CG - #ATTCTATA        6480                                                                           - TTTCATCATA ACAGTAGTAC ATTAATCAGT GATATACTGA AACGATCTAC AG - #ACTCAACT        6540                                                                           - ATGCAAGGAA TAAGCAATAT GCCAATTATG TCTAATATTT TAACTTTAGA AC - #TAAAACGT        6600                                                                           - TCTACCAATA CTAAAAATAG GATACGTGAT AGGCTGTTAA AAGCTGCAAT AA - #ATAGTAAG        6660                                                                           - GATGTAGAAG AAATACTTTG TTCTATACCT TCGGAGGAAA GAACTTTAGA AC - #AACTTAAG        6720                                                                           #          6749    TTTA TGAAGGTAC                                              - (2) INFORMATION FOR SEQ ID NO:178:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 5798 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:178:                                - TCTAGAACTA GTGGATCTTC TGGTAATGAC AAATTAAACT GTTTAGCGTA TA - #TTATATAC          60                                                                           - TCGTATAAAA AATCATGATC TATATTCTTA ATAGCTTTTA GAAGGTTCAT AT - #CGTAGAAA         120                                                                           - TAAACATAAG TTCCTTTCAT CACTCTACCT ACACGACCTT TACGTTGCGT CA - #TCATAGAT         180                                                                           - TTTGATATAA ACATCTGAAC ACCACCAAAA GGTCTAGGTA CGTATACTCT AC - #CGGTATCG         240                                                                           - TATACGTGAG TCGCTGTACG TATAGTAATA CTAGATTCCA AATAAGGGGT AG - #ATACTAGA         300                                                                           - ATACAAGGTC TTTCTCTATT AGGTCGTTGA ACATCTTGTA GGATTTCTGC TA - #TATTTTTT         360                                                                           - AATTTTCCAT GTATTACTAT AAAATCAATA TTCTTATTCT TAGATTCTAA GT - #ACTCTTTA         420                                                                           - TACTTAATAC ATTCTGATAC AGAAGGTAAG AATAAAATAC CACACATTCC AT - #TATCTGGC         480                                                                           - TTACACCACA ATAAAGTAGA CGATATATTC TTTCTCTCAT TATCAAAATA AA - #CTCTCTTA         540                                                                           - TCCGGAGAAT ACCTATTTTT TACGTATATT TCTTTTATGG AGTAAAGAAC TG - #GTCCTTCT         600                                                                           - ATATGGTAAA ATTCAACATC AGGAAGAAAT TCCATTAGTC TATCTTTATC AT - #CTTCTAGA         660                                                                           - GTGGCAGACA TCAATACTAG CGAATGAATG CTATCTATAT TTTTTCTTAG AA - #CGGCTATC         720                                                                           - ATAATATCGG CTATCCTATC ATGTTCATGT ATTTCATCTA TTATGACTAT AT - #TATACTTT         780                                                                           - GATAGAGAGT AACTAGTCAG TTTATTAGTA GAAAGTACTA TACCTTGAAA TC - #CTTTTTTG         840                                                                           - GTTTTTTCTG TATGTCCTCC GTATTTAAGT TCTACAGGAG AACCTTCGAA CT - #GTGAAAAT         900                                                                           - CCCAACGATT GTAAAAAATT ATTTCCGTTG CTCTTTACCA AAGTCACCCT AG - #GAAGAGAT         960                                                                           - AAAACTATAG GTTTGGGTAT AAAATCTAGC CTTATCCTGT CTATATCATC CC - #ATCCTCCG        1020                                                                           - AATAAATAGT TATACCACAT TATTACTTTT GGTAACTGAG ATGTTTTACC TA - #TGCCTGTA        1080                                                                           - CTACCGGTAA CTACTATCTG TTTCCTCTTC TTTAACATAT CAAAGATATG AA - #CCTGTGTT        1140                                                                           - GTTAAACTAA GGGATTTGAA CGATATGATA GCGAAAGGAT TTGGATTATT GA - #GTATTCCT        1200                                                                           - ATAGAATTCT TAATGGGTAC CTTCTTATTG GAAGAGAAAA TAGACAGATG AT - #TTCCAGCT        1260                                                                           - ACTAGTAATC CTCTTTTATC GTCAAGCGTT ATATCAGATA CATGATTATA AC - #CGATACAT        1320                                                                           - TTTACGTAAC TATAGCATTC AAACGTTATA AATCTATCGT TACCTATATA GT - #ATACCTGT        1380                                                                           - TTACTGTAGT TGATACTGAC GGGTATTATA TCTATAAGTT TACTAACAGG TA - #TTTTAGCG        1440                                                                           - GGTATTGAAT TAGTAGTTTC TATATTCAGC ATATAAGTAT CGTCCTTTAA GC - #AGATAAAT        1500                                                                           - ACTTTATTCC ACCTATGTTT TATTATAGGA AATACAGAAT GAGAAAAAAA TA - #ACGTATCT        1560                                                                           - TTATTATGAT ATTCTTCTAA TTCTTTTTGG GTATACTTAC TTGGGAATAT AT - #CGTACATA        1620                                                                           - TTAGGGAAAG CGTATATCGA AAATAGCTCG TTAGTGGCCA TAGTTCCTAC AG - #TATGTATA        1680                                                                           - TTTAGTTAGT AATAAATGGA TAGATACACA GAACTAGTTA TTAATAAAAT AC - #CAGAATTA        1740                                                                           - GGATTCGTTA ACTTGCTTTC TCATATCTAT CAAACAGTTG GGTTATCCTA CG - #ATATAGAT        1800                                                                           - GTATCAAAAT TCAAAACTAA TTGCAATGGT TACGTCGTAG AGAGATTTGA TA - #ACTCAGAA        1860                                                                           - ACAGTTGGCA AAGTGTCCTG CGTGCCTATA TCTATACTGT TAGAATTGGT AG - #ACAGAAAA        1920                                                                           - ATATTATCTA AACCAGATAC GTCTAAAACA GAAATAGAGA TTAAAGAAGA TT - #TAGTAAAC        1980                                                                           - GAATTAATTG AAAATACCAA TAGTTTCGAA GATATAATGA CTATACCTAC CA - #GTATCCCT        2040                                                                           - ATGAGATATT TTTTTAAACC GGTACTAAGA GAAAAAGTAT CTAAAGCTGT AG - #ATTTTTCC        2100                                                                           - AGAATGGATA TTAAGGGAGA TGATATTAGC AAAATGGGAA TAAAACACGG AG - #AAAAAAGT        2160                                                                           - AATAATATAT CTAATATTAA GATTGTACCA GAAAAAGATG CCTGGATGAC TA - #ATACTAGT        2220                                                                           - ATTCAGCAAT TAATAGGACC TATGTCGTAC GGAACAGAAG TTAGCTATAT AG - #GTCAATTT        2280                                                                           - AACTTTAATT TTATTAACAC ATATCCTGTA TACGAAAAAT CTGCAGCCCT TA - #ACAGAAGT        2340                                                                           - CCAGAACTTT TTAAGATTAA AGATAGAATT AAAGGATTAC GTACAAGATT TG - #TTATGTTC        2400                                                                           - GGTTTCTGTT ATATGTTCCA TTGGAAATGT TTGATATATG ATAGAGAAAA CG - #ATTTTGTA        2460                                                                           - TGTTTCTATG ATTCAGGAGG ATCTAATCCA AATGACTTTG ATCACTATGA TA - #ATTTTTTC        2520                                                                           - TACTATAGTC ATTCGAGAGG ATTCAATAGA AATTCTAAGA GGTCATCTAG CT - #TATCTAAT        2580                                                                           - GAAAATGCAG ATATAGATAT TCTGTTCAAC TTTTTCGTGG ATAATTACGA AG - #TTACTTCA        2640                                                                           - GGATGTATAA ACGTAGAAGT CAATCAGCTG ATGGAATCAG AATGTGGTAT GT - #TTACTTGT        2700                                                                           - TTGTTTATGA CTATGTGCTG TCTCCATCCT CCTAAAGGAT TTAAAGGGAT AA - #GAAAGACA        2760                                                                           - TATACCTATT TTAAGTTTTT AGCCGATAAA AAAATGACTA TGCTAAAGTC TA - #TACTTTTC        2820                                                                           - AACGCTGACA AGATGGAATT TAAAGTGAAA GAATCAAGCA GTAAAGGCAT AC - #AAGAATAT        2880                                                                           - AAAAAAATGG AAGAGTGGTG TGGTAAAACT ATAAACATTT TAGCTGATAA AA - #TAACAACA        2940                                                                           - CGTGTAAATA GTATAATAGA GTAGTAAAAT GGATAATTTT ATAAAGCAGA TA - #TCGTCAAA        3000                                                                           - GATAGTAAAA CCTATAGCAG AATTAGAACC TCCAGATTCT AAAGTACAAT AT - #TATTACAT        3060                                                                           - GACTATATCG TTTAATTTTC CTGACTTATA TTATTGTAAT AAAAATTTAT TT - #GCGAAACC        3120                                                                           - CGATAATACT TTGCTAGATG TTTCTAAGTC TTTGCTTACT TTAAACTCAT TT - #CCGTATGA        3180                                                                           - AAACTTTGTG ATAAATGATT TACTAAGAAC TATTAGGCGT TACTGTCACG TA - #TATGATGT        3240                                                                           - CTATTTTTTA CCCGTAGGGT GGTTTGTAGG AAAAGAAGAT GTATTACCCA AT - #TACCAAGT        3300                                                                           - ATCGATAAAA ATAATAAGAA GTACTAATCA AGAAGTAATA GAAAACATTA TT - #AGGAATTA        3360                                                                           - TTTATCACGA CACGGTATTT ATGGAGATAA CCTATCTATA GAAACAGACC GA - #TTAAACGA        3420                                                                           - AGTATCTATA AACAGACATT CTATTGTAGG AGCTAGACAG TTAGCACCTA TA - #TGCGTTGT        3480                                                                           - TTCTTTTTAT CCTTTCGACC CTGAAAATAA AATACTTTTC GTTATATATG TA - #GGTAGATA        3540                                                                           - CAAAGACAGA CATTGCGGTG TATCTTATGT AGTTGATAGA GAGGATATGT AT - #AAAGTAAT        3600                                                                           - TAACAGAATA TATTCTTACG TAGTTTGTAT TTATCTAGTT TCCGATGATA TG - #GTCACGTT        3660                                                                           - TCATACTACT CCTCTAGCTA ATCACAGTAA AAAATTAATA CCGTTACCCA TA - #AATCATTG        3720                                                                           - CAATACCTTA TGCGAGATAG TTCACGACTT TGAGTTTTTA AGATTTGAGC AA - #TCCACTAT        3780                                                                           - GCCAATACCC GTTTTCACTC CTTTTATTCC TAAACAGCTA GTTAATATAA TC - #AACTTACC        3840                                                                           - TGATGATATA CCTATTACTT GTGCATCAAT AAACAGATTA GAATATGTTA CA - #CATATAGA        3900                                                                           - TGATAAAAAA TTAAAAAGAG TACTGATTAT CGTAAAGGAT AAATTTCTTA GA - #AATACTAT        3960                                                                           - TCTTCACGGT ACATTTAAAA AAAGGAATAT AGTCAGAAAC AGGAAATATA CT - #TTCACTAT        4020                                                                           - AACATGGTCT AATTTCGAAT GTCCGACGTT AGGAGACGTT AAGTCTTCTT CA - #CCTAATAC        4080                                                                           - CTGTAATAGA GTAGTTTTAG ACGGTAGTAG ATACGTTACA AAAACCTTTA AT - #GATACAAT        4140                                                                           - ATAAATGGAA CTAACTAGAG AAACGCTGAT ATTTGTAGGC ATTACTGTAC TA - #GTAGTAGT        4200                                                                           - AATGATCATA TCTGGTTTCT CACTAATATT GCGATTGATA CCTGGTGTAT AT - #TCATCAGT        4260                                                                           - TATTAGATCG TCGTTCGTAG GAGGGAAAAT ATTAAGATTT ATGGAGGTAT TC - #TCTACTGT        4320                                                                           - TATGTTTATA CCATCATTAG TAATACTTTA TACAGCATAT ATAAGGAAAT CT - #AAAGTGAA        4380                                                                           - AAATAACTAA ATATTATAGT ATTTGTAATA AATGGCTACT GGAGAGATTC GT - #CTTATTAT        4440                                                                           - AGGGCCTATG TTTTCAGGTA AAACAACAGA ATTAGTTAGA TTAATAAGAA GA - #TTTATGAT        4500                                                                           - ATCGGGACGT AAATGTATAA TAATAAAACA TTGTAGTGAT TCCCGTTATA CC - #GAAGGAGA        4560                                                                           - TTTAGAAGCT ATATATACTC ATGATAAAAT TTCGATGGAA GCACTATCGT GT - #AGCAAATT        4620                                                                           - ATTACCTTTA ATACCTAAAA TTGATAACTT TGAAGTAATA GGTATAGACG AA - #GGACAGTT        4680                                                                           - TTTTGAAGAT ATAGTAGAAT TTAGTGAGAT TATGGCTAAT AAGGGTAAAA CT - #GTAATCAT        4740                                                                           - AGCGGCTTTA AATGGAGATT TCAAACGACA ATTATTTGGA AACATATTTA AA - #CTATTATC        4800                                                                           - TTTATCAGAA TCAGTTACTA GTTTAACTGC TATTTGTGCA GTTTGTAAAA AC - #GAAGCATC        4860                                                                           - TTTTTCTAAG CGCATGACTG ATGATAAAGA TGTAAAAGTT ATAGGAGGTA AA - #GAAATGTA        4920                                                                           - TACTGCTGTT TGTAGAAAAT GCTTTTTATG AGTCTAATAT ACGTACTAAA TA - #CTTGTACG        4980                                                                           - TACAACTATG TTAGAATAAT TTGCTTAGTA TAGTATATAA ACAAGTATGT AA - #AAAATAAA        5040                                                                           - ATTGATATAA AAGTAGTCTT CTATTCCGAA CAATAACTAT ACAAAATGGA TT - #TAGATATT        5100                                                                           - AAATCTTGCA GAAGTATTTA CAAAATATGG GATAAATATC ATTTTATGAC AG - #GGTATAAA        5160                                                                           - TATAAAAATG ATAAACAGAG ATTTAAAATT ACAATTTACT GTAAATGTGA TT - #GTTCTATC        5220                                                                           - AAAGAATATC CTTATAGATT TGTTACTGAG AAACTGCTTT TAATGTATAT TA - #TTAATAAG        5280                                                                           - TTTAGAGGAA AGTATCTAAT CAAAATTAGG ATAGAACCCA TAGTTAAAAA TT - #AAATCATA        5340                                                                           - TATCAATACA TGTCAGTTTT TTATCGAAAA ATGGATTTAT AAATAAAATG AA - #AAATAACT        5400                                                                           - TGAATGAAGG AAAAAATAAC CATGAGTAAA AAACCAGTAA AGACGGTCCA GC - #GTAGACGT        5460                                                                           - GGAAACGATG AGGATAATAA GTTTACTTGT ATCCAAGCGC TAGAACATGC AA - #AAAGCTTA        5520                                                                           - TGTACTAAAA ATAATAAAAT AGTTAAATCT GTTAAACTAT CACAATCTCT CT - #TTAAGTCA        5580                                                                           - TCTAACAATA TTTCTGTGAT ATTAGAACCA GAATATAAAG ACAAATTAGT GA - #CTCCTCTT        5640                                                                           - ATTATTGTAG AAGGTGAAGG AAAAATATAC CATAATAAGA ATGATAGTTT TA - #ATCGTGAA        5700                                                                           - GAACCGTATT TTCTAAAAAT ACGACCTACG TTAATGAATC CTATATTATA TC - #AGATTATG        5760                                                                           #   5798           ATCC CCCGGGCTGC AGGAATTC                                    - (2) INFORMATION FOR SEQ ID NO:179:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:179:                                # 20               CATG                                                        - (2) INFORMATION FOR SEQ ID NO:180:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 70 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:180:                                - TATGAATTCC TCGAGGGATC CAGGCCTTTT TTATTGACTA GTTAATCAGT CT - #AATATACG          60                                                                           #        70                                                                    - (2) INFORMATION FOR SEQ ID NO:181:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 20 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:181:                                # 20               GACG                                                        - (2) INFORMATION FOR SEQ ID NO:182:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 66 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:182:                                - TTAGAATTCT CGCGACCCGG GTTTTTATAG CTAATTAGTA CTTATTACAA AT - #ACTATAAT          60                                                                           #           66                                                                 - (2) INFORMATION FOR SEQ ID NO:183:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 35 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:183:                                #       35         CCTC GAGGGTACCG CATGC                                       - (2) INFORMATION FOR SEQ ID NO:184:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 31 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:184:                                #          31      AGGG ATCCGTCGAC G                                           - (2) INFORMATION FOR SEQ ID NO:185:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 52 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:185:                                - CCGAAGCTTC TCGAGATAAA AATCAACGAC TGTCGGTAGC GTCCACGACG AC - #                  52                                                                           - (2) INFORMATION FOR SEQ ID NO:186:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 16 base                                                            (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:186:                                #    16                                                                        - (2) INFORMATION FOR SEQ ID NO:187:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 5062 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:187:                                - AAGCTTGCGG CCGCTCATTA GACAAGCGAA TGAGGGACGA AAACGTGGAG GA - #GGTATTAA          60                                                                           - GTTTGGAGAA ATGGAGAGAG ACTGTTTAAT AGCGCATGGC GCAGCCAATA CT - #ATTACAGA         120                                                                           - AGTTTTGAAA GATTCGGAAG AAGATTATCA AGATGTGTAT GTTTGTGAAA AT - #TGTGGAGA         180                                                                           - CATAGCAGCA CAAATCAAGG GTATTAATAC ATGTCTTAGA TGTTCAAAAC TT - #AATCTCTC         240                                                                           - TCCTCTCTTA ACAAAAATTG ATACCACGCA CGTATCTAAA GTATTTCTTA CT - #CAAATGAA         300                                                                           - CGCCAGAGGC GTAAAAGTCA AATTAGATTT CGAACGAAGG CCTCCTTCGT TT - #TATAAACC         360                                                                           - ATTAGATAAA GTTGATCTCA AGCCGTCTTT TCTGGTGTAA TAAAAATTAA TT - #AATTACTC         420                                                                           - GAGATAAAAA TCAACGACTG TCGGTAGCGT CCACGACGAC GTCAGTTACT TC - #TAGGACCG         480                                                                           - TACCGTTTTT CAAAAGCATG AGGTAGTGAG TTCGCGGAGA TGAGACCACC AC - #TTCGTTGT         540                                                                           - AGGGATCCAG GGCGAAAAGG ACGTCGTCCG AGTCGTGCAT GTACATGATG TT - #GATGACGC         600                                                                           - CTTGCGTGTC GTCGTATTCT AGTAGGGCGC TTTGGCAAAA GGCGCAGTTT TC - #TAGGGAAA         660                                                                           - TGTTGAGCGC CGCTGTGATG CTGTGTGTGG TATGCATGTT GCGCGTCAGT TC - #GCATTTAG         720                                                                           - TTTGACTGTC CGTCTGGGTG ATGATGAGGC TCTGGCCTAC GACGGTGGTG GA - #GACAGGGT         780                                                                           - AGGAGATACC TTTGATCAGG TACTGGTTTG TTACGACATA ACTGACGTGT TC - #GGAGACGG         840                                                                           - TCAGCGCGGA GAAGGATTCG CCGAGCGGCA GACAAAACAG GTCGGGGAAG GT - #TTCTAGCG         900                                                                           - TGCTTGGTTG CATGGTAGAT AGGATGGAGA GGGCGGCGGG AACGGTAGTG GG - #GACGGTGG         960                                                                           - CATCGGGGAA GAGACGTGTG AGGCGTTCGA GCGAGTGATC GCGTCGCCCG CT - #ACTGGAAC        1020                                                                           - AGGGTGTGTA CAGGTCGCTG AGGTATTCGT GGTGCGGATG AGCTAGCAAC TG - #CGTAAAGT        1080                                                                           - GTGATAGCTC GGCTAATGAA CAGAGGCCCG TTTCTACGAT GAAGATTTCG CG - #TCTCTCCG        1140                                                                           - TCGTATGTAC TAGCATGGAG TGGACGAGGC TGCCCATGAG GTAGAGTTCT TG - #ACGCGCGA        1200                                                                           - AGGCTGAAAG AAAAGAGGCC AGGTGCGTTT TGTGTAGTTT TAGGGCAAAG TC - #GGCGATCT        1260                                                                           - GTCGTAGTGC CCACTGGGGG ATGAGATGTT GCTGATTCTG TTTAGAGAGT AT - #GTAGACCA        1320                                                                           - GGCGTACGAG GCTGGTGATG TCGGTGATCT GATTCGGTGT CCAAAGGGCT CG - #TTTGGCCA        1380                                                                           - GGTCCACGGC CGTGGGATAC AGCAGCAACG TGGTGCGTGG TGGTGTTTGT GA - #GAGGCAGG        1440                                                                           - TGATCATAAA TTCTTGTATT TGTAAGAGTG CGGCCTGGCG GTCTAGGGCC CG - #TGGGACGG        1500                                                                           - AGACTTGGGC GCCGGCCTCT TCTTGTCGGG CTGCTGCGAA CAGTGCTAAT GC - #GTAGGCGA        1560                                                                           - AGGCCATTTC TACCGTGCGG CGGTCCAGCA TCTGACATCG ACCGCTTTTG AG - #TACATCCA        1620                                                                           - CGGCGTAACG GTGAAAGCTG TTACGTAGTA GTGCGCTGAG GTCCAGGTAG TT - #GAAGTCAA        1680                                                                           - GTGCGGCGTC AAGAAAGTCC GGGTCTTTGA GATAAGAGTG ACGGTTCAGT TG - #ATCTTTCT        1740                                                                           - TAACTAGCAC CAGGAGCTCG TGTTTTTCAG TTTGTCGTAG TATAAAGTTG TC - #GCGTTGAT        1800                                                                           - AGGGCGCTTT AAAGAGTACG CGTGGAAGAT GGCCGAAGAT AAGCAGCATG GG - #TGTGTCGT        1860                                                                           - CGTCTATGGA CACCGTAACT ACGAAGAAGT CCTCGGTCAG TGTTATTTTA AC - #GTAACGTA        1920                                                                           - GTTCGTCGAT GAGGTAAAAG CCTTGGTGCA AACAAGGTGT GACGGTGCTG AA - #TAGTAGAT        1980                                                                           - CGTGTCCATC AAAGAGGATA CAGGTCTGGT TAAAGTGTGG TCGGTGTAGT CC - #TGAGGTGG        2040                                                                           - TATGTGATTC TGTCCAGCCG TGTGGAGTGG TTTGCGGTGG CATCCAAACG TG - #AGGTATTG        2100                                                                           - ACAGGTCAAT GGGTGGTGGC ACAGTGGTGG GCTGTTCACC TAGGCTGTCC TG - #TGCCTTTA        2160                                                                           - GCTGCTGCGA AAAAGATCGG TAGCTGGCCA GGTCTTTGGA TACCAGCGCG TA - #AGTGTTAA        2220                                                                           - GTCTCTGTTG GTATCTTTCC AGGGTTTCGG TCAGATCTAC CTGGTTCAGA AA - #CTGCTCCG        2280                                                                           - CCAGAGGACC CGCAAAAAGA CATCGAGGCA TATGGAATAC ATAGTATTGA TT - #ATAGCTTT        2340                                                                           - GGAAAAAGTT GAAACTGATG GCGTTTTCCC TGACGACCGT GCTGTTACGG AG - #GCTGCTAT        2400                                                                           - TGTAGGTACA CTGGGTGGTG TTTTCACGCA GGAAGCGGAT GGGTCTCCCG TA - #GGTGTTGA        2460                                                                           - GCAGTAGGTG AAACGCTTTG TCCAGCGGTT CGGATATGGC TTCTGCGCCA TA - #TCGTGACG        2520                                                                           - AAAGTAGGTG GCTGAGGAGA CAGACGGCGA GGACGATGAG GTAGGAGGGG AG - #CCCGGGCC        2580                                                                           - GCATTTTATA TTGTAATTAT ATATTTTCAA TTTTGAAATC CCAAAATATT AT - #CATATTCT        2640                                                                           - TCCCAATAAA CTCGAGGGTA CCGGATCCTT CTTTATTCTA TACTTAAAAA GT - #GAAAATAA        2700                                                                           - ATACAAAGGT TCTTGAGGGT TGTGTTAAAT TGAAAGCGAG AAATAATCAT AA - #ATTATTTC        2760                                                                           - ATTATCGCGA TATCCGTTAA GTTTGTATCG TAATGTGCCG CCGCCCGGAT TG - #CGGCTTCT        2820                                                                           - CTTTCTCACC TGGACCGGTG GCACTGCTGT GGTGTTGCCT TCTGCTGCCC AT - #CGTTTCCT        2880                                                                           - CAGCCACCGT CAGCGTCGCT CCTACCGTCG CCGAGAAAGT TCCCGCGGAG TG - #CCCCGAAC        2940                                                                           - TAACGCGTCG ATGCCTGTTG GGTGAGGTGT TTCAGGGTGA CAAGTATGAA AG - #TTGGCTGC        3000                                                                           - GCCCGTTGGT GAATGTTACC AGACGCGATG GCCCGCTATC GCAACTTATT CG - #TTACCGTC        3060                                                                           - CCGTTACGCC GGAGGCCGCC AACTCCGTGC TGTTGGACGA TGCTTTCCTG GA - #CACTCTGG        3120                                                                           - CCCTGCTGTA CAACAATCCG GATCAATTGC GGGCCTTGCT GACGCTGTTG AG - #CTCGGACA        3180                                                                           - CAGCGCCGCG CTGGATGACG GTGATGCGCG GTTACAGCGA GTGCGGCGAT GG - #CTCGCCGG        3240                                                                           - CCGTGTACAC GTGCGTGGAC GACCTGTGCC GCGGCTACGA CCTCACGCGA CT - #GTCATACG        3300                                                                           - GGCGCAGCAT CTTCACGGAA CACGTGTTAG GCTTCGAGCT GGTGCCACCG TC - #TCTCTTTA        3360                                                                           - ACGTGGTGGT GGCCATACGC AACGAAGCCA CGCGTACCAA CCGCGCCGTG CG - #TCTGCCCG        3420                                                                           - TGAGCACCGC TGCCGCGCCC GAGGGCATCA CGCTCTTTTA CGGCCTGTAC AA - #CGCAGTGA        3480                                                                           - AGGAATTCTG CCTGCGTCAC CAGCTGGACC CGCCGCTGCT ACGCCACCTA GA - #TAAATACT        3540                                                                           - ACGCCGGACT GCCGCCCGAG CTGAAGCAGA CGCGCGTCAA CCTGCCGGCT CA - #CTCGCGCT        3600                                                                           - ATGGCCCTCA AGCAGTGGAT GCTCGCTAAT TTTTATAGAT CCCCCGGGAA TC - #GATTCGCG        3660                                                                           - ATAGCTGATT AGTTTTTGTT AACAAAAATG TGGGAGAATC TAATTAGTTT TT - #CTTTACAC        3720                                                                           - AATTGACGTA CATGAGTCTG AGTTCCTTGT TTTTGCTAAT TATTTCATCC AA - #TTTATTAT        3780                                                                           - TCTTGACGAT ATCGAGATCT TTTGTATAGG AGTCAGACTT GTATTCAACA TG - #CTTTTCTA        3840                                                                           - TAATCATCTT AGTTATTTCG GCATCATCCA ATAGTACATT TTCCAGATTA AC - #AGAGTAGA        3900                                                                           - TATTAATGTC GTATTTGAAC AGAGCCTGTA ACATCTCAAT GTCTTTATTA TC - #TATAGCCA        3960                                                                           - ATTTAATGTC CGGAATGAAG AGAAGGGAAT TATTGGTGTT TGTCGACGTC AT - #ATAGTCGA        4020                                                                           - GCAAGAGAAT CATCATATCC ACGTGTCCAT TTTTTATAGT GGTGTGAATA CA - #ACTAAGGA        4080                                                                           - GAATAGCCAG ATCAAAAGTA GATGGTATTT CTGAAAGAAA GTATGATACA AT - #ACTTACAT        4140                                                                           - CATTAAGCAT GACGGCATGA TAAAATGAAG TTTTCCATCC AGTTTTCCCA TA - #GAACATCA        4200                                                                           - GTCTCCAATT TTTCTTAAAC AGTTTCACCG TTTGCATGTT ACCACTATCA AC - #CGCATAAT        4260                                                                           - ACAATGCGGT GTTTCCTTTG TCATCAAATT GTGAATCATC CATTCCACTG AA - #TAGCAAAA        4320                                                                           - TCTTTACTAT TTTGGTATCT TCTAATGTGG CTGCCTGATG TAATGGAAAT TC - #ATTCTCTA        4380                                                                           - GAAGATTTTT CAATGCTCCA GCGTTCAACA ACGTACATAC TAGACGCACG TT - #ATTATCAG        4440                                                                           - CTATTGCATA ATACAAGGCA CTATGTCCAT GGACATCCGC CTTAAATGTA TC - #TTTACTAG        4500                                                                           - AGAGAAAGCT TTTCAGCTGC TTAGACTTCC AAGTATTAAT TCGTGACAGA TC - #CATGTCTG        4560                                                                           - AAACGAGACG CTAATTAGTG TATATTTTTT CATTTTTTAT AATTTTGTCA TA - #TTGCACCA        4620                                                                           - GAATTAATAA TATCTCTAAT AGATCTAATT TAATTTAATT TATATAACTT AT - #TTTTTGAA        4680                                                                           - TATACTTTTA ATTAACAAAA GAGTTAAGTT ACTCATATGG ACGCCGTCCA GT - #CTGAACAT        4740                                                                           - CAATCTTTTT AGCCAGAGAT ATCATAGCCG CTCTTAGAGT TTCAGCGTGA TT - #TTCCAACC        4800                                                                           - TAAATAGAAC TTCATCGTTG CGTTTACAAC ACTTTTCTAT TTGTTCAAAC TT - #TGTTGTTA        4860                                                                           - CATTAGTAAT CTTTTTTTCC AAATTAGTTA GCCGTTGTTT GAGAGTTTCC TC - #ATTGTCGT        4920                                                                           - CTTCATCGGC TTTAACAATT GCTTCGCGTT TAGCCTCCTG GCTGTTCTTA TC - #AGCCTTTG        4980                                                                           - TAGAAAAAAA TTCAGTTGCT GGAATTGCAA GATCGTCATC TCCGGGGAAA AG - #AGTTCCGT        5040                                                                           #               5062AAT TC                                                     - (2) INFORMATION FOR SEQ ID NO:188:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 2151 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:188:                                - ATGCGGCCAG GCCTCCCCTC CTACCTCATC GTCCTCGCCG TCTGTCTCCT CA - #GCCACCTA          60                                                                           - CTTTCGTCAC GATATGGCGC AGAAGCCATA TCCGAACCGC TGGACAAAGC GT - #TTCACCTA         120                                                                           - CTGCTCAACA CCTACGGGAG ACCCATCCGC TTCCTGCGTG AAAACACCAC CC - #AGTGTACC         180                                                                           - TACAATAGCA GCCTCCGTAA CAGCACGGTC GTCAGGGAAA ACGCCATCAG TT - #TCAACTTT         240                                                                           - TTCCAAAGCT ATAATCAATA CTATGTATTC CATATGCCTC GATGTCTTTT TG - #CGGGTCCT         300                                                                           - CTGGCGGAGC AGTTTCTGAA CCAGGTAGAT CTGACCGAAA CCCTGGAAAG AT - #ACCAACAG         360                                                                           - AGACTTAACA CTTACGCGCT GGTATCCAAA GACCTGGCCA GCTACCGATC TT - #TTTCGCAG         420                                                                           - CAGCTAAAGG CACAGGACAG CCTAGGTGAA CAGCCCACCA CTGTGCCACC AC - #CCATTGAC         480                                                                           - CTGTCAATAC CTCACGTTTG GATGCCACCG CAAACCACTC CACACGGCTG GA - #CAGAATCA         540                                                                           - CATACCACCT CAGGACTACA CCGACCACAC TTTAACCAGA CCTGTATCCT CT - #TTGATGGA         600                                                                           - CACGATCTAC TATTCAGCAC CGTCACACCT TGTTTGCACC AAGGCTTTTA CC - #TCATCGAC         660                                                                           - GAACTACGTT ACGTTAAAAT AACACTGACC GAGGACTTCT TCGTAGTTAC GG - #TGTCCATA         720                                                                           - GACGACGACA CACCCATGCT GCTTATCTTC GGCCATCTTC CACGCGTACT CT - #TTAAAGCG         780                                                                           - CCCTATCAAC GCGACAACTT TATACTACGA CAAACTGAAA AACACGAGCT CC - #TGGTGCTA         840                                                                           - GTTAAGAAAG ATCAACTGAA CCGTCACTCT TATCTCAAAG ACCCGGACTT TC - #TTGACGCC         900                                                                           - GCACTTGACT TCAACTACCT GGACCTCAGC GCACTACTAC GTAACAGCTT TC - #ACCGTTAC         960                                                                           - GCCGTGGATG TACTCAAAAG CGGTCGATGT CAGATGCTGG ACCGCCGCAC GG - #TAGAAATG        1020                                                                           - GCCTTCGCCT ACGCATTAGC ACTGTTCGCA GCAGCCCGAC AAGAAGAGGC CG - #GCGCCCAA        1080                                                                           - GTCTCCGTCC CACGGGCCCT AGACCGCCAG GCCGCACTCT TACAAATACA AG - #AATTTATG        1140                                                                           - ATCACCTGCC TCTCACAAAC ACCACCACGC ACCACGTTGC TGCTGTATCC CA - #CGGCCGTG        1200                                                                           - GACCTGGCCA AACGAGCCCT TTGGACACCG AATCAGATCA CCGACATCAC CA - #GCCTCGTA        1260                                                                           - CGCCTGGTCT ACATACTCTC TAAACAGAAT CAGCAACATC TCATCCCCCA GT - #GGGCACTA        1320                                                                           - CGACAGATCG CCGACTTTGC CCTAAAACTA CACAAAACGC ACCTGGCCTC TT - #TTCTTTCA        1380                                                                           - GCCTTCGCGC GTCAAGAACT CTACCTCATG GGCAGCCTCG TCCACTCCAT GC - #TAGTACAT        1440                                                                           - ACGACGGAGA GACGCGAAAT CTTCATCGTA GAAACGGGCC TCTGTTCATT AG - #CCGAGCTA        1500                                                                           - TCACACTTTA CGCAGTTGCT AGCTCATCCG CACCACGAAT ACCTCAGCGA CC - #TGTACACA        1560                                                                           - CCCTGTTCCA GTAGCGGGCG ACGCGATCAC TCGCTCGAAC GCCTCACACG TC - #TCTTCCCC        1620                                                                           - GATGCCACCG TCCCCACTAC CGTTCCCGCC GCCCTCTCCA TCCTATCTAC CA - #TGCAACCA        1680                                                                           - AGCACGCTAG AAACCTTCCC CGACCTGTTT TGTCTGCCGC TCGGCGAATC CT - #TCTCCGCG        1740                                                                           - CTGACCGTCT CCGAACACGT CAGTTATGTC GTAACAAACC AGTACCTGAT CA - #AAGGTATC        1800                                                                           - TCCTACCCTG TCTCCACCAC CGTCGTAGGC CAGAGCCTCA TCATCACCCA GA - #CGGACAGT        1860                                                                           - CAAACTAAAT GCGAACTGAC GCGCAACATG CATACCACAC ACAGCATCAC AG - #CGGCGCTC        1920                                                                           - AACATTTCCC TAGAAAACTG CGCCTTTTGC CAAAGCGCCC TACTAGAATA CG - #ACGACACG        1980                                                                           - CAAGGCGTCA TCAACATCAT GTACATGCAC GACTCGGACG ACGTCCTTTT CG - #CCCTGGAT        2040                                                                           - CCCTACAACG AAGTGGTGGT CTCATCTCCG CGAACTCACT ACCTCATGCT TT - #TGAAAAAC        2100                                                                           #           2151AAGTAAC TGACGTCGTC GTGGACGCTA CCGACAGTCG T                     - (2) INFORMATION FOR SEQ ID NO:189:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 3962 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:189:                                - GCCCTTAACA GAAGTCCAGA ACTTTTTAAG ATTAAAGATA GAATTAAAGG AT - #TACGTACA          60                                                                           - AGATTTGTTA TGTTCGGTTT CTGTTATATG TTCCATTGGA AATGTTTGAT AT - #ATGATAGA         120                                                                           - GAAAACGATT TTGTATGTTT CTATGATTCA GGAGGATCTA ATCCAAATGA CT - #TTGATCAC         180                                                                           - TATGATAATT TTTTCTACTA TAGTCATTCG AGAGGATTCA ATAGAAATTC TA - #AGAGGTCA         240                                                                           - TCTAGCTTAT CTAATGAAAA TGCAGATATA GATATTCTGT TCAACTTTTT CG - #TGGATAAT         300                                                                           - TACGAAGTTA CTTCAGGATG TATAAACGTA GAAGTCAATC AGCTGATGGA AT - #CAGAATGT         360                                                                           - GGTATGTTTA CTTGTTTGTT TATGACTATG TGCTGTCTCC ATCCTCCTAA AG - #GATTTAAA         420                                                                           - GGGATAAGAA AGACATATAC CTATTTTAAG TTTTTAGCCG ATAAAAAAAT GA - #CTATGCTA         480                                                                           - AAGTCTATAC TTTTCAACGC TGACAAGATG GAATTTAAAG TGAAAGAATC AA - #GCAGTAAA         540                                                                           - GGCATACAAG AATATAAAAA AATGGAAGAG TGGTGTGGTA AAACTATAAA CA - #TTTTAGCT         600                                                                           - GATAAAATAA CAACACGTGT AAATAGTATA ATAGAGTAGT AAAATGGATA AT - #TTTATAAA         660                                                                           - GCAGATATCG TCAAAGATAG TAAAACCTAT AGCAGAATTA GAACCTCCAG AT - #TCTAAAGT         720                                                                           - ACAATATTAT TACATGACTA TATCGTTTAA TTTTCCTGAC TTATATTATT GT - #AATAAAAA         780                                                                           - TTTATTTGCG AAACCCGATA ATACTTTGCT AGATGTTTCT AAGTCTTTGC TT - #ACTTTAAA         840                                                                           - CTCATTTCCG TATGAAAACT TTGTGATAAA TGATTTACTA AGAACTATTA GG - #CGTTACTG         900                                                                           - TCACGTATAT GATGTCTATT TTTTACCCGT AGGTGGTTTG TAGGAAAAGA AG - #ATGTATTA         960                                                                           - CCCAATTACC AAGTATCGAT AAAAATAATA AGAAGTACTA ATCAAGAAGT AA - #TAGAAAAC        1020                                                                           - ATTATTAGGA ATTATTTATC ACGACACGGT ATTTATGGAG ATAACCTATC TA - #TAGAAACA        1080                                                                           - GACCGATTAA ACGAAGTATC TATAAACAGA CATTCTATTG TAGGAGCTAG AC - #AGTTAGCA        1140                                                                           - CCTATATGCG TTGTTTCTTT TTATCCTTTC GACCCTGAAA ATAAAATACT TT - #TCGTTATA        1200                                                                           - TATGTAGGTA GATACAAAGA CAGACATTGC GGTGTATCTT ATGTAGTTGA TA - #GAGAGGAT        1260                                                                           - ATGTATAAAG TAATTAACAG AATATATTCT TACGTAGTTT GTATTTATCT AG - #TTTCCGAT        1320                                                                           - GATATGGTCA CGTTTCATAC TACTCCTCTA GCTAATCACA GTAAAAAATT AA - #TACCGTTA        1380                                                                           - CCCATAAATC ATTGCAATAC CTTATGCGAG ATAGTTCACG ACTTTGAGTT TT - #TGAGATTT        1440                                                                           - GAGCAATCCA CTATGCCAAT ACCCGTTTTC ACTCCTTTTA TTCCTAAACA GC - #TAGTTAAT        1500                                                                           - ATAATCAACT TACCTGATGA TATACCTATT ACTTGTGCAT CAATAAACAG AT - #TAGAATAT        1560                                                                           - GTTACACATA TAGATGATAA AAAATTAAAA AGAGTACTGA TTATCGTAAA GG - #ATAAATTT        1620                                                                           - CTTAGAAATA CTATTCTTCA CGGTACATTT AAAAAAAGGA ATATAGTCAG AA - #ACAGGAAA        1680                                                                           - TATACTTTCA CTATAACATG GTCTAATTTC GAATGTCCGA CGTTAGGAGA CG - #TTAAGTCT        1740                                                                           - TCTTCACCTA ATACCTGTAA TAGAGTAGTT TTAGACGGTA GTAGATACGT TA - #CAAAAACC        1800                                                                           - TTTAATGATA CAATATAAAT GGAACTAACT AGAGAAACGC TGATATTTGT AG - #GCATTACT        1860                                                                           - GTACTAGTAG TAGTAATGAT CATATCTGGT TTCTCACTAA TATTGCGATT GA - #TACCTGGT        1920                                                                           - GTATATTCAT CAGTTATTAG ATCGTCGTTC GTAGGAGGGA AAATATTAAG AT - #TTATGGAG        1980                                                                           - GTATTCTCTA CTGTTATGTT TATACCATCA TTAGTAATAC TTTATACAGC AT - #ATATAAGG        2040                                                                           - AAATCTAAAG TGAAAAATAA CTAAATATTA TAGTATTTGT AATAAGTACT AA - #TTAGCTAT        2100                                                                           - AAAAACCCGG GTCGCGAGAA TTCGTCGACG GATCCTTCTT TATTCTATAC TT - #AAAAAGTG        2160                                                                           - AAAATAAATA CAAAGGTTCT TGAGGGTTGT GTTAAATTGA AAGCGAGAAA TA - #ATCATAAA        2220                                                                           - TTATTTCATT ATCGCGATAT CCGTTAAGTT TGTATCGTAA TGTGCCGCCG CC - #CGGATTGC        2280                                                                           - GGCTTCTCTT TCTCACCTGG ACCGGTGGCA CTGCTGTGGT GTTGCCTTCT GC - #TGCCCATC        2340                                                                           - GTTTCCTCAG CCACCGTCAG CGTCGCTCCT ACCGTCGCCG AGAAAGTTCC CG - #CGGAGTGC        2400                                                                           - CCCGAACTAA CGCGTCGATG CCTGTTGGGT GAGGTGTTTC AGGGTGACAA GT - #ATGAAAGT        2460                                                                           - TGGCTGCGCC CGTTGGTGAA TGTTACCAGA CGCGATGGCC CGCTATCGCA AC - #TTATTCGT        2520                                                                           - TACCGTCCCG TTACGCCGGA GGCCGCCAAC TCCGTGCTGT TGGACGATGC TT - #TCCTGGAC        2580                                                                           - ACTCTGGCCC TGCTGTACAA CAATCCGGAT CAATTGCGGG CCTTGCTGAC GC - #TGTTGAGC        2640                                                                           - TCGGACACAG CGCCGCGCTG GATGACGGTG ATGCGCGGTT ACAGCGAGTG CG - #GCGATGGC        2700                                                                           - TCGCCGGCCG TGTACACGTG CGTGGACGAC CTGTGCCGCG GCTACGACCT CA - #CGCGACTG        2760                                                                           - TCATACGGGC GCAGCATCTT CACGGAACAC GTGTTAGGCT TCGAGCTGGT GC - #CACCGTCT        2820                                                                           - CTCTTTAACG TGGTGGTGGC CATACGCAAC GAAGCCACGC GTACCAACCG CG - #CCGTGCGT        2880                                                                           - CTGCCCGTGA GCACCGCTGC CGCGCCCGAG GGCATCACGC TCTTTTACGG CC - #TGTACAAC        2940                                                                           - GCAGTGAAGG AATTCTGCCT GCGTCACCAG CTGGACCCGC CGCTGCTACG CC - #ACCTAGAT        3000                                                                           - AAATACTACG CCGGACTGCC GCCCGAGCTG AAGCAGACGC GCGTCAACCT GC - #CGGCTCAC        3060                                                                           - TCGCGCTATG GCCCTCAAGC AGTGGATGCT CGCTAATTTT TATAGATCCC TC - #GAGGGTAC        3120                                                                           - CGCATGCCCT TTTTATTGAC TAGTTAATCA GTCTAATATA CGTACTAAAT AC - #TTGTACGT        3180                                                                           - ACAACTATGT TAGAATAATT TGCTTAGTAT AGTATATAAA CAAGTATGTA AA - #AAATAAAA        3240                                                                           - TTGATATAAA AGTAGTCTTC TATTCCGAAC AATAACTATA CAAAATGGAT TT - #AGATATTA        3300                                                                           - AATCTTGCAG AAGTATTTAC AAAATATGGG ATAAATATCA TTTTATGACA GG - #GTATAAAT        3360                                                                           - ATAAAAATGA TAAACAGAGA TTTAAAATTA CAATTTACTG TAAATGTGAT TG - #TTCTATCA        3420                                                                           - AAGAATATCC TTATAGATTT GTTACTGAGA AACTGCTTTT AATGTATATT AT - #TAATAAGT        3480                                                                           - TTAGAGGAAA GTATCTAATC AAAATTAGGA TAGAACCCAT AGTTAAAAAT TA - #AATCATAT        3540                                                                           - ATCAATACAT GTCAGTTTTT TATCGAAAAA TGGATTTATA AATAAAATGA AA - #AATAACTT        3600                                                                           - GAATGAAGGA AAAAATAACC ATGAGTAAAA AACCAGTAAA GACGGTCCAG CG - #TAGACGTG        3660                                                                           - GAAACGATGA GGATAATAAG TTTACTTGTA TCCAAGCGCT AGAACATGCA AA - #AAGCTTAT        3720                                                                           - GTACTAAAAA TAATAAAATA GTTAAATCTG TTAAACTATC ACAATCTCTC TT - #TAAGTCAT        3780                                                                           - CTAACAATAT TTCTGTGATA TTAGAACCAG AATATAAAGA CAAATTAGTG AC - #TCCTCTTA        3840                                                                           - TTATTGTAGA AGGTGAAGGA AAAATATACC ATAATAAGAA TGATAGTTTT AA - #TCGTGAAG        3900                                                                           - AACCGTATTT TCTAAAAATA CGACCTACGT TAATGAATCC TATATTATAT CA - #GATTATGG        3960                                                                           #            3962                                                              - (2) INFORMATION FOR SEQ ID NO:190:                                           -      (i) SEQUENCE CHARACTERISTICS:                                           #pairs    (A) LENGTH: 5302 base                                                          (B) TYPE: nucleic acid                                                         (C) STRANDEDNESS: single                                                       (D) TOPOLOGY: linear                                                 -     (ii) MOLECULE TYPE: DNA (genomic)                                        -     (xi) SEQUENCE DESCRIPTION: SEQ ID NO:190:                                - GCCCTTAACA GAAGTCCAGA ACTTTTTAAG ATTAAAGATA GAATTAAAGG AT - #TACGTACA          60                                                                           - AGATTTGTTA TGTTCGGTTT CTGTTATATG TTCCATTGGA AATGTTTGAT AT - #ATGATAGA         120                                                                           - GAAAACGATT TTGTATGTTT CTATGATTCA GGAGGATCTA ATCCAAATGA CT - #TTGATCAC         180                                                                           - TATGATAATT TTTTCTACTA TAGTCATTCG AGAGGATTCA ATAGAAATTC TA - #AGAGGTCA         240                                                                           - TCTAGCTTAT CTAATGAAAA TGCAGATATA GATATTCTGT TCAACTTTTT CG - #TGGATAAT         300                                                                           - TACGAAGTTA CTTCAGGATG TATAAACGTA GAAGTCAATC AGCTGATGGA AT - #CAGAATGT         360                                                                           - GGTATGTTTA CTTGTTTGTT TATGACTATG TGCTGTCTCC ATCCTCCTAA AG - #GATTTAAA         420                                                                           - GGGATAAGAA AGACATATAC CTATTTTAAG TTTTTAGCCG ATAAAAAAAT GA - #CTATGCTA         480                                                                           - AAGTCTATAC TTTTCAACGC TGACAAGATG GAATTTAAAG TGAAAGAATC AA - #GCAGTAAA         540                                                                           - GGCATACAAG AATATAAAAA AATGGAAGAG TGGTGTGGTA AAACTATAAA CA - #TTTTAGCT         600                                                                           - GATAAAATAA CAACACGTGT AAATAGTATA ATAGAGTAGT AAAATGGATA AT - #TTTATAAA         660                                                                           - GCAGATATCG TCAAAGATAG TAAAACCTAT AGCAGAATTA GAACCTCCAG AT - #TCTAAAGT         720                                                                           - ACAATATTAT TACATGACTA TATCGTTTAA TTTTCCTGAC TTATATTATT GT - #AATAAAAA         780                                                                           - TTTATTTGCG AAACCCGATA ATACTTTGCT AGATGTTTCT AAGTCTTTGC TT - #ACTTTAAA         840                                                                           - CTCATTTCCG TATGAAAACT TTGTGATAAA TGATTTACTA AGAACTATTA GG - #CGTTACTG         900                                                                           - TCACGTATAT GATGTCTATT TTTTACCCGT AGGTGGTTTG TAGGAAAAGA AG - #ATGTATTA         960                                                                           - CCCAATTACC AAGTATCGAT AAAAATAATA AGAAGTACTA ATCAAGAAGT AA - #TAGAAAAC        1020                                                                           - ATTATTAGGA ATTATTTATC ACGACACGGT ATTTATGGAG ATAACCTATC TA - #TAGAAACA        1080                                                                           - GACCGATTAA ACGAAGTATC TATAAACAGA CATTCTATTG TAGGAGCTAG AC - #AGTTAGCA        1140                                                                           - CCTATATGCG TTGTTTCTTT TTATCCTTTC GACCCTGAAA ATAAAATACT TT - #TCGTTATA        1200                                                                           - TATGTAGGTA GATACAAAGA CAGACATTGC GGTGTATCTT ATGTAGTTGA TA - #GAGAGGAT        1260                                                                           - ATGTATAAAG TAATTAACAG AATATATTCT TACGTAGTTT GTATTTATCT AG - #TTTCCGAT        1320                                                                           - GATATGGTCA CGTTTCATAC TACTCCTCTA GCTAATCACA GTAAAAAATT AA - #TACCGTTA        1380                                                                           - CCCATAAATC ATTGCAATAC CTTATGCGAG ATAGTTCACG ACTTTGAGTT TT - #TGAGATTT        1440                                                                           - GAGCAATCCA CTATGCCAAT ACCCGTTTTC ACTCCTTTTA TTCCTAAACA GC - #TAGTTAAT        1500                                                                           - ATAATCAACT TACCTGATGA TATACCTATT ACTTGTGCAT CAATAAACAG AT - #TAGAATAT        1560                                                                           - GTTACACATA TAGATGATAA AAAATTAAAA AGAGTACTGA TTATCGTAAA GG - #ATAAATTT        1620                                                                           - CTTAGAAATA CTATTCTTCA CGGTACATTT AAAAAAAGGA ATATAGTCAG AA - #ACAGGAAA        1680                                                                           - TATACTTTCA CTATAACATG GTCTAATTTC GAATGTCCGA CGTTAGGAGA CG - #TTAAGTCT        1740                                                                           - TCTTCACCTA ATACCTGTAA TAGAGTAGTT TTAGACGGTA GTAGATACGT TA - #CAAAAACC        1800                                                                           - TTTAATGATA CAATATAAAT GGAACTAACT AGAGAAACGC TGATATTTGT AG - #GCATTACT        1860                                                                           - GTACTAGTAG TAGTAATGAT CATATCTGGT TTCTCACTAA TATTGCGATT GA - #TACCTGGT        1920                                                                           - GTATATTCAT CAGTTATTAG ATCGTCGTTC GTAGGAGGGA AAATATTAAG AT - #TTATGGAG        1980                                                                           - GTATTCTCTA CTGTTATGTT TATACCATCA TTAGTAATAC TTTATACAGC AT - #ATATAAGG        2040                                                                           - AAATCTAAAG TGAAAAATAA CTAAATATTA TAGTATTTGT AATAAGTACT AA - #TTAGCTAT        2100                                                                           - AAAAACCCGG GCTCGAGATA AAAATTACTG GTCAGCCTTG CTTCTAGTCA CC - #ATAGGGTG        2160                                                                           - GGTACTCTTA CCTCCAGAGG CGGTGGGTTC CTCAGCACCA TCCTCCTCTT CC - #TCTGGGGC        2220                                                                           - AACTTCCTCT ATCTCAGACA CTGGCTCAGA CTTGACAGAC ACAGTGTCCT CC - #CGCTCCTC        2280                                                                           - CTGAGCACCC TCCTCCTCTT CCTCATCACT CTGCTCACTT TCTTCCTGAT CA - #CTGTTCTC        2340                                                                           - AGCCACAATT ACTGAGGACA GAGGGATAGT CGCGGGTACA GGGGACTCTG GG - #GGTGACAC        2400                                                                           - CAGAGAATCA GAGGAGCTGA CACCAGCGGT GGCCAAAGTG TAGGCTACAA TA - #GCCTCTTC        2460                                                                           - CTCATCTGAC TCCTCGGCGA TGGCCCGTAG GTCATCCACA CTAGGAGAGC AG - #ACTCTCAG        2520                                                                           - AGGATCGGCC CCCAGAATGT ACTGGGCAAA GACCTTCATG CAGATCTCCT CA - #ATGCGGCG        2580                                                                           - CTTCATTACA CTGATAACCT CAGGCTTGGT TATCAGAGGC CGCTTGGCCA GC - #ATCACACT        2640                                                                           - AGTCTCCTCT AAGACATAGC AGCACAGCAC CCGACAGAAC TCACTTAAGA GA - #GAGATGCC        2700                                                                           - CCCGTACATG GTCATCATAC AAGCGTCACT AGTGACCTTG TACTCATTAC AC - #ATTGTTTC        2760                                                                           - CACACATGTA GTGAGGATAT CCATAAATAT GTGATCAATG TGCGTGAGCA CC - #TTGTCTCT        2820                                                                           - CTCCTCATCC AAAATCTTAA ATATTTTCTG GGCATAAGCC ATAATCTCAT CA - #GGGGAGCA        2880                                                                           - CTGAGGCAAG TTCTGCAGTG CCGCCATGGC CTGACTGCAG CCATTGGTGG TC - #TTAGGGAA        2940                                                                           - GGCTGAGTTC TTGGTAAAGA ACTCTATATT CCTGTAGCAC ATATACATCA TC - #TTTCTCCT        3000                                                                           - AAGTTCATCC TTTTTAGCAC GGGCCTTAGC CTGCAGTGCA CCCCCCAACT TG - #TTAGCGGC        3060                                                                           - GCCCTTGCTC ACATCATGCA GCTCCTTAAT ACAAGCCATC CACATCTCCC GC - #TTATCCTC        3120                                                                           - AGGTACAATG TAGTTCTCAT ACATGCTCTG CATAGTTAGC CCAATACACT TC - #ATCTCCTC        3180                                                                           - GAAAGGCTCA TGAACCTTAT CTAAGATATC TAAGGCATTC TGCAAACATC CT - #CCCATCAT        3240                                                                           - ATTAAAGGCG CCAGTGAATT TCTCTTCCGT CTGGGTATAT TTTTTCAGCA TG - #TGCTCCTT        3300                                                                           - GATTCTATGC CGCACCATGT CCACTCGAAC CTTAATCTGT TTCATTACGA TA - #CAAACTTA        3360                                                                           - ACGGATATCG CGATAATGAA ATAATTTATG ATTATTTCTC GCTTTCAATT TA - #ACACAACC        3420                                                                           - CTCAAGAACC TTTGTATTTA TTTTCACTTT TTAAGTATAG AATAAAGAAG GA - #TCCTTCTT        3480                                                                           - TATTCTATAC TTAAAAAGTG AAAATAAATA CAAAGGTTCT TGAGGGTTGT GT - #TAAATTGA        3540                                                                           - AAGCGAGAAA TAATCATAAA TTATTTCATT ATCGCGATAT CCGTTAAGTT TG - #TATCGTAA        3600                                                                           - TGTGCCGCCG CCCGGATTGC GGCTTCTCTT TCTCACCTGG ACCGGTGGCA CT - #GCTGTGGT        3660                                                                           - GTTGCCTTCT GCTGCCCATC GTTTCCTCAG CCACCGTCAG CGTCGCTCCT AC - #CGTCGCCG        3720                                                                           - AGAAAGTTCC CGCGGAGTGC CCCGAACTAA CGCGTCGATG CCTGTTGGGT GA - #GGTGTTTC        3780                                                                           - AGGGTGACAA GTATGAAAGT TGGCTGCGCC CGTTGGTGAA TGTTACCAGA CG - #CGATGGCC        3840                                                                           - CGCTATCGCA ACTTATTCGT TACCGTCCCG TTACGCCGGA GGCCGCCAAC TC - #CGTGCTGT        3900                                                                           - TGGACGATGC TTTCCTGGAC ACTCTGGCCC TGCTGTACAA CAATCCGGAT CA - #ATTGCGGG        3960                                                                           - CCTTGCTGAC GCTGTTGAGC TCGGACACAG CGCCGCGCTG GATGACGGTG AT - #GCGCGGTT        4020                                                                           - ACAGCGAGTG CGGCGATGGC TCGCCGGCCG TGTACACGTG CGTGGACGAC CT - #GTGCCGCG        4080                                                                           - GCTACGACCT CACGCGACTG TCATACGGGC GCAGCATCTT CACGGAACAC GT - #GTTAGGCT        4140                                                                           - TCGAGCTGGT GCCACCGTCT CTCTTTAACG TGGTGGTGGC CATACGCAAC GA - #AGCCACGC        4200                                                                           - GTACCAACCG CGCCGTGCGT CTGCCCGTGA GCACCGCTGC CGCGCCCGAG GG - #CATCACGC        4260                                                                           - TCTTTTACGG CCTGTACAAC GCAGTGAAGG AATTCTGCCT GCGTCACCAG CT - #GGACCCGC        4320                                                                           - CGCTGCTACG CCACCTAGAT AAATACTACG CCGGACTGCC GCCCGAGCTG AA - #GCAGACGC        4380                                                                           - GCGTCAACCT GCCGGCTCAC TCGCGCTATG GCCCTCAAGC AGTGGATGCT CG - #CTAATTTT        4440                                                                           - TATAGATCCC TCGAGGGTAC CGCATGCCCT TTTTATTGAC TAGTTAATCA GT - #CTAATATA        4500                                                                           - CGTACTAAAT ACTTGTACGT ACAACTATGT TAGAATAATT TGCTTAGTAT AG - #TATATAAA        4560                                                                           - CAAGTATGTA AAAAATAAAA TTGATATAAA AGTAGTCTTC TATTCCGAAC AA - #TAACTATA        4620                                                                           - CAAAATGGAT TTAGATATTA AATCTTGCAG AAGTATTTAC AAAATATGGG AT - #AAATATCA        4680                                                                           - TTTTATGACA GGGTATAAAT ATAAAAATGA TAAACAGAGA TTTAAAATTA CA - #ATTTACTG        4740                                                                           - TAAATGTGAT TGTTCTATCA AAGAATATCC TTATAGATTT GTTACTGAGA AA - #CTGCTTTT        4800                                                                           - AATGTATATT ATTAATAAGT TTAGAGGAAA GTATCTAATC AAAATTAGGA TA - #GAACCCAT        4860                                                                           - AGTTAAAAAT TAAATCATAT ATCAATACAT GTCAGTTTTT TATCGAAAAA TG - #GATTTATA        4920                                                                           - AATAAAATGA AAAATAACTT GAATGAAGGA AAAAATAACC ATGAGTAAAA AA - #CCAGTAAA        4980                                                                           - GACGGTCCAG CGTAGACGTG GAAACGATGA GGATAATAAG TTTACTTGTA TC - #CAAGCGCT        5040                                                                           - AGAACATGCA AAAAGCTTAT GTACTAAAAA TAATAAAATA GTTAAATCTG TT - #AAACTATC        5100                                                                           - ACAATCTCTC TTTAAGTCAT CTAACAATAT TTCTGTGATA TTAGAACCAG AA - #TATAAAGA        5160                                                                           - CAAATTAGTG ACTCCTCTTA TTATTGTAGA AGGTGAAGGA AAAATATACC AT - #AATAAGAA        5220                                                                           - TGATAGTTTT AATCGTGAAG AACCGTATTT TCTAAAAATA CGACCTACGT TA - #ATGAATCC        5280                                                                           #               5302TGG AA                                                     __________________________________________________________________________ 

What is claimed is:
 1. A recombinant poxvirus containing therein exogenous DNA from HCMV coding for an HCMV protein selected from the group consisting of gB, gH and combinations thereof, wherein the poxvirus is selected from the group consisting of:(i) recombinant vaccinia virus wherein regions C7L-K1L, J2R, B13R+B14R, A26L A56R and I4L have been deleted therefrom, or wherein the open reading frames for the thymidine kinase gene, the hemorrhagic region, the A type inclusion body region, the hemagglutinin gene, the host range gene region, and the large subunit, ribonucleotide reductase have been deleted therefrom; (ii) NYVAC vaccinia virus; and (iii) ALVAC canarypox virus.
 2. The recombinant poxvirus of claim 1 wherein J2R, B13R+B14R, A26L, A56R, C7L-K1L and 14L are deleted from the virus.
 3. The recombinant poxvirus of claim 1 wherein a thymidine kinase gene, a hemorrhagic region, an A type inclusion body region, a hemagglutinin gene, a host range region, and a large subunit, ribonucleotide reductase are deleted from the virus.
 4. The recombinant poxvirus of claim 1 which is a NYVAC recombinant virus.
 5. The recombinant poxvirus of claim 1 which is an ALVAC recombinant virus.
 6. The recombinant poxvirus of claim 1 which is vCP233, vP1360, ALVAC-CMV6, ALVAC-CMV5, vCP236 or vCP139.
 7. The recombinant poxvirus of claim 1 which is vP1173, vP1183, vP1312, vP1302B, vP1399, or vP1001.
 8. A method for treating a patient in need of immunological treatment or of inducing an immunological response in an individual or animal comprising administering to said patient or individual or animal a composition comprising a virus as claimed in any one of claims 1, 2, 3, 4 or 5 in admixture with a suitable carrier.
 9. A composition for inducing an immunological response comprising a virus as claimed in any one of claims 1, 2, 3, 4 or 5 in admixture with a suitable carrier.
 10. A method for expressing a gene product in a cell cultured in vitro comprising introducing into the cell a virus as claimed in any one of claims 1, 2, 3, 4 or 5 in admixture with a suitable carrier.
 11. The method of claim 8 further comprising administering an HCMV antigen either before or after administering the composition.
 12. The method of claim 11 wherein the antigen is from the in vitro expression of a recombinant avipox virus or vaccinia virus. 